Balázs Harrach - Academia.edu (original) (raw)

Papers by Balázs Harrach

Research paper thumbnail of Molecular characterization of a lizard adenovirus reveals the first atadenovirus with two fiber genes and the first adenovirus with either one short or three long fibers per penton

Journal of virology, 2014

Although adenoviruses (AdVs) have been found in a wide variety of reptiles, including numerous sq... more Although adenoviruses (AdVs) have been found in a wide variety of reptiles, including numerous squamate species, turtles, and crocodiles, the number of reptilian adenovirus isolates is still scarce. The only fully sequenced reptilian adenovirus, snake adenovirus 1 (SnAdV-1), belongs to the Atadenovirus genus. Recently, two new atadenoviruses were isolated from a captive Gila monster (Heloderma suspectum) and Mexican beaded lizards (Heloderma horridum). Here we report the full genomic and proteomic characterization of the latter, designated lizard adenovirus 2 (LAdV-2). The double-stranded DNA (dsDNA) genome of LAdV-2 is 32,965 bp long, with an average G+C content of 44.16%. The overall arrangement and gene content of the LAdV-2 genome were largely concordant with those in other atadenoviruses, except for four novel open reading frames (ORFs) at the right end of the genome. Phylogeny reconstructions and plesiomorphic traits shared with SnAdV-1 further supported the assignment of LAdV...

Research paper thumbnail of DNA sequence of a small, unidentified plasmid isolated from a Haemophilus somnus strain: Short communication

Acta Veterinaria Hungarica

One of the plasmids present in a Haemophilus somnus strain isolated from nasal discharge of a cat... more One of the plasmids present in a Haemophilus somnus strain isolated from nasal discharge of a cattle with respiratory disease was purified and cloned for DNA sequencing. The plasmid was found to be 1065 base pairs long with 39.2% G + C content, and showed no homology to any DNA sequenced so far. It has no capacity to code any protein longer than 43 residues. It is not clear yet if this plasmid codes Haemophilus somnus specific factors.

Research paper thumbnail of Introduction of the family Alloherpesviridae: the first molecular detection of herpesviruses of cyprinid fish in Hungary

Magyar Allatorvosok Lapja

The taxonomy of herpesviruses has been changed recently radically. In 2008, a new order, the Herp... more The taxonomy of herpesviruses has been changed recently radically. In 2008, a new order, the Herpesvirales was established to contain the family Herpesviridae along with two additional new families, Alloherpesviridae and Malacoherpesviridae. The family Alloherpesviridae includes the herpesviruses isolated from amphibians and fish, whereas Malacoherpesviridae contains the single herpesvirus originating from an invertebrate host, namely an oyster. Previously, these viruses had been classified into the family Herpesviridae based on their characteristic morphology, although they did not fit unambiguously into any genus or subfamily of the family. The newly established family Alloherpesviridae is now officially divided into four genera. One of these contains the herpesviruses of amphibians, whereas the other three contain those of different fishes. The genus Cyprinivirus includes three virus species originating from cyprinid fishes. Cyprinid herpesvirus 1 (CyHV-1) is the causative agent ...

Research paper thumbnail of Crystal structure of the fibre head domain of bovine adenovirus 4, a ruminant atadenovirus

Virology journal, 2015

In adenoviruses, primary host cell recognition is generally performed by the head domains of thei... more In adenoviruses, primary host cell recognition is generally performed by the head domains of their homo-trimeric fibre proteins. This first interaction is reversible. A secondary, irreversible interaction subsequently takes place via other adenovirus capsid proteins and leads to a productive infection. Although many fibre head structures are known for human mastadenoviruses, not many animal adenovirus fibre head structures have been determined, especially not from those belonging to adenovirus genera other than Mastadenovirus. We constructed an expression vector for the fibre head domain from a ruminant atadenovirus, bovine adenovirus 4 (BAdV-4), consisting of amino acids 414-535, expressed the protein in Escherichia coli, purified it by metal affinity and cation exchange chromatography and crystallized it. The structure was solved using single isomorphous replacement plus anomalous dispersion of a mercury derivative and refined against native data that extended to 1.2 Å resolution....

Research paper thumbnail of Infectious canine hepatitis: detection of canine adenovirus type 1 by polymerase chain reaction

Acta veterinaria Hungarica, 1996

A primer pair and a polymerase chain reaction (PCR) method earlier tested for the detection of hu... more A primer pair and a polymerase chain reaction (PCR) method earlier tested for the detection of human and animal adenoviruses were used to demonstrate the presence of canine adenovirus type 1 (CAV-1) in tissue culture and clinical specimens. A simple procedure of sample preparation was elaborated making the PCR easily applicable in rapid confirmation of the diagnosis of infectious canine hepatitis.

Research paper thumbnail of Four new inverted terminal repeat sequences from bovine adenoviruses reveal striking differences in the length and content of the ITRs

Virus genes, 2001

The inverted terminal repeat (ITR) of the genome of four bovine adenovirus (BAdV) types have been... more The inverted terminal repeat (ITR) of the genome of four bovine adenovirus (BAdV) types have been sequenced, analysed and compared to the ITRs of other adenoviruses. The length of ITRs of the examined BAdVs ranged between 59 and 368 base pairs, thus the presently known longest adenovirus ITR sequence is from BAdV-10. The conserved motifs and characteristic sequence elements of the ITRs providing different binding sites for replicative proteins of viral and cellular origin seemed to be distributed according to the proposed genus classification of BAdVs. The ITRs of BAdV-10 share similarity with the members of the genus Mastadenovirus, while the ITRs of the other three sequenced serotypes (BAdV-4, 5 and strain Rus) which are candidate members of the genus Atadenovirus are very short and contain NFI and Sp1 binding sites only. The analysis of the new ITRs implied that the nucleotide sequence of the so-called core origin is highly preserved within the mastadenovirus genus only.

Research paper thumbnail of Restriction site mapping of bovine adenovirus type 1

Acta veterinaria Hungarica, 1990

Physical maps of the DNA of bovine adenovirus (BAV) type 1 were established using ApaI, BamHI, Bs... more Physical maps of the DNA of bovine adenovirus (BAV) type 1 were established using ApaI, BamHI, BstEII, EcoRI and KpnI enzymes. The size of the viral genome was found to be around 35,000 base pairs. The orientation of the maps was determined by hybridizing Southern blots of BAV-1 DNA with the cloned hexon gene region of BAV-3.

Research paper thumbnail of Siadenovirus

The Springer Index of Viruses, 2002

Research paper thumbnail of Sequence, Transcriptional Analysis, and Deletion of the Bovine Adenovirus Type 1 E3 Region

Virology, 1998

The early 3 (E3) transcriptional unit of human adenoviruses (HAV) encodes proteins that modulate ... more The early 3 (E3) transcriptional unit of human adenoviruses (HAV) encodes proteins that modulate host antiviral immune defenses. HAV E3 sequences are highly variable; different HAV groups encode phylogenetically unrelated proteins. The role of the E3 region of many human and animal adenoviruses is unknown because the sequences are unrelated to previously characterized viruses and the functions of proteins encoded by these regions have not been studied. We sequenced a portion of the bovine adenovirus serotype 1 (BAV-1) genome corresponding to the putative E3 region. This sequence was substantially different from other adenoviral E3 sequences, including those of two other bovine adenoviruses. However, two regions of putative sequence conservation were identified. BAV-1 E3 sequences were identified in early and late transcripts, but, unlike HAV, introns were not detected in the E3 region transcripts. Like HAV E3, a majority of the BAV-1 E3 region was not essential for growth in cell culture, as demonstrated by the construction of a recombinant BAV-1 lacking 60% of the putative E3 region.

Research paper thumbnail of Genomic characterization of human adenovirus 36, a putative obesity agent

Virus Research, 2010

Increased levels of serum antibody titers against human adenovirus 36 (HAdV-D36) are associated w... more Increased levels of serum antibody titers against human adenovirus 36 (HAdV-D36) are associated with human obesity and experimental obesity in laboratory animals. While HAdV-D36 has been studied as an infectious agent implicated in obesity for over a decade, the complete genome sequence and its analysis have yet to be reported. A detailed analysis of the genome sequence of HAdV-D36 may be important to understand its role in obesity. Genomic and bioinformatic comparisons with other HAdVs identified differences that suggested unique functions. Global pairwise genome alignment with all sequenced human adenovirus D (HAdV-D) genomes revealed areas of nonconserved sequences in the hexon, E3 CR1␤, E3 CR1␥, and fiber genes. Phylogenetic analysis of all HAdV-D36 proteins confirmed that this virus belongs to species Human adenovirus D. This genomic analysis of HAdV-D36 provides an important tool for comprehending the role that this unique adenovirus may play in human obesity. Low amino acid sequence identity in the E3 CR1␤ and CR1␥ genes may suggest distinctive roles for these proteins. Furthermore, the predicted molecular models of the HAdV-D36 fiber protein seem to implicate a unique tissue tropism for HAdV-D36.

Research paper thumbnail of Molecular characterization of a lizard adenovirus reveals the first atadenovirus with two fiber genes and the first adenovirus with either one short or three long fibers per penton

Journal of virology, 2014

Although adenoviruses (AdVs) have been found in a wide variety of reptiles, including numerous sq... more Although adenoviruses (AdVs) have been found in a wide variety of reptiles, including numerous squamate species, turtles, and crocodiles, the number of reptilian adenovirus isolates is still scarce. The only fully sequenced reptilian adenovirus, snake adenovirus 1 (SnAdV-1), belongs to the Atadenovirus genus. Recently, two new atadenoviruses were isolated from a captive Gila monster (Heloderma suspectum) and Mexican beaded lizards (Heloderma horridum). Here we report the full genomic and proteomic characterization of the latter, designated lizard adenovirus 2 (LAdV-2). The double-stranded DNA (dsDNA) genome of LAdV-2 is 32,965 bp long, with an average G+C content of 44.16%. The overall arrangement and gene content of the LAdV-2 genome were largely concordant with those in other atadenoviruses, except for four novel open reading frames (ORFs) at the right end of the genome. Phylogeny reconstructions and plesiomorphic traits shared with SnAdV-1 further supported the assignment of LAdV...

Research paper thumbnail of Sequencing and phylogenetic analysis of the protease gene, and genetic mapping of bovine adenovirus type 10 define its relatedness to other bovine adenoviruses

Virus Research, 1998

The complete genome of a bovine adenovirus (BAV) type 10 isolate was molecularly cloned and parti... more The complete genome of a bovine adenovirus (BAV) type 10 isolate was molecularly cloned and partially sequenced. The encoded proteins were predicted by computer analysis of the DNA sequences of the ends or the entire length of the cloned viral fragments, and thus a rough genetic map was constructed. The protease gene of BAV-10 was completely sequenced and used in phylogenetic analysis. Based on the results of the phylogenetic analysis, and the location and presence of certain genes thought to be specifically characteristic of subgroup 1 or subgroup 2 BAVs, it could be concluded that, in spite of the striking similarity in certain biological properties, BAV-10 is not related to subgroup 2 BAVs as originally described. It does not however fit clearly into subgroup 1 either, the members of which show closer relationship with human adenoviruses. BAV-10 therefore should best be considered as the first member of a third subgroup of BAVs.

Research paper thumbnail of A novel siadenovirus detected in the kidneys and liver of Gouldian finches (Erythura gouldiae)

Veterinary Microbiology, 2014

This article appeared in a journal published by Elsevier. The attached copy is furnished to the a... more This article appeared in a journal published by Elsevier. The attached copy is furnished to the author for internal non-commercial research and education use, including for instruction at the authors institution and sharing with colleagues.

Research paper thumbnail of Molecular typing of fowl adenoviruses, isolated in Hungary recently, reveals high diversity

Veterinary Microbiology, 2013

Molecular typing of 26 recent fowl adenovirus (FAdV) isolates obtained in Eastern Hungary between... more Molecular typing of 26 recent fowl adenovirus (FAdV) isolates obtained in Eastern Hungary between 2006 and 2011 was performed. The viruses were isolated on primary chicken embryo liver cell cultures from carcasses of chickens, sent for diagnostic investigation. The isolates were subjected to a PCR to amplify a fragment from the hexon gene. Sequence analysis of the amplicons revealed the presence of all the five FAdV species in the country, among them FAdV-B, which has been barely found in Hungary before. The three strains classified as FAdV-B might represent a novel type within this species, as their nucleotide sequence identity to strain 340 (type FAdV-5) is below the commonly accepted intratype limit. Most of the strains (63%) were classified into species FAdV-E (12 strains) and FAdV-D (7 strains) in the study. Four of the isolates proved to be the mixture of two adenovirus strains belonging to two different FAdV types, in three from these four cases, these two types even belonged to two different species. Analysing the pathological findings bolsters certain established connections between some FAdV types and disease conditions. Gizzard erosion was found in connection with FAdV-1 strains only, and inclusion body hepatitis with FAdV-D and -E strains.

Research paper thumbnail of Detection and Analysis of Six Lizard Adenoviruses by Consensus Primer PCR Provides Further Evidence of a Reptilian Origin for the Atadenoviruses

Journal of Virology, 2004

A consensus nested-PCR method was designed for investigation of the DNA polymerase gene of adenov... more A consensus nested-PCR method was designed for investigation of the DNA polymerase gene of adenoviruses. Gene fragments were amplified and sequenced from six novel adenoviruses from seven lizard species, including four species from which adenoviruses had not previously been reported. Host species included Gila monster, leopard gecko, fat-tail gecko, blue-tongued skink, Tokay gecko, bearded dragon, and mountain chameleon. This is the first sequence information from lizard adenoviruses. Phylogenetic analysis indicated that these viruses belong to the genus Atadenovirus, supporting the reptilian origin of atadenoviruses. This PCR method may be useful for obtaining templates for initial sequencing of novel adenoviruses.

Research paper thumbnail of Recognition and partial genome characterization by non-specific DNA amplification and PCR of a new siadenovirus species in a sample originating from Parus major, a great tit

Journal of Virological Methods, 2010

A seemingly novel siadenovirus species was detected by PCR and sequencing in the sample of a grea... more A seemingly novel siadenovirus species was detected by PCR and sequencing in the sample of a great tit (Parus major) found dead in Hungary. Since the genus Siadenovirus has very few known members so far, further study of the virus was intriguing not only from epizootiological but also from taxonomical aspects. The sample, which had been tested in another PCR survey previously, consisted of less than 50 l of extracted nucleic acid. To ensure sufficient target DNA for an extended study, the viral genome had to be preserved. To this end, the sample was subjected to a novel method of non-specific DNA amplification. Using the amplified DNA as target, different PCR and sequencing strategies were applied with consensus or specific primers for the study of the central genome part of the putative tit adenovirus. The sequence of supposedly one half (13,628 bp) of the genome was determined including eight full genes between the genes of the IVa2 and hexon proteins. The gene content of the viral genome fragment as well as the results of the phylogenetic analyses with different proteins confirmed the discovery of a new species in the genus Siadenovirus. This is the first report on the detection of an adenovirus in great tits. The methods, described in this work, proved suitable for the recovery of nucleic acid samples that contain irreplaceable microbial genomic DNA but are only available in limited quantities.

Research paper thumbnail of DNA sequencing and analysis of the right-hand part of the genome of the unique bovine adenovirus type 10

Journal of General Virology, 2004

DNA sequencing and analysis of the right-hand part of the genome of the unique bovine adenovirus ... more DNA sequencing and analysis of the right-hand part of the genome of the unique bovine adenovirus type 10

Research paper thumbnail of Genome sequence of a waterfowl aviadenovirus, goose adenovirus 4

Journal of General Virology, 2012

We present, to our knowledge, the first complete genome sequence of a waterfowl aviadenovirus, go... more We present, to our knowledge, the first complete genome sequence of a waterfowl aviadenovirus, goose adenovirus (GoAdV) strain P29, and an analysis of its genetic content in comparison with five published aviadenovirus genome sequences. Of the 35 genes predicted to encode functional proteins, the central region of the genome contains 19 (IVa2 to fiber-2) that were inherited from the ancestor of all known adenoviruses. Of the remaining genes, nine have orthologues only in aviadenoviruses and seven lack orthologues in any adenovirus. We also obtained limited sequence data for a pathogenic GoAdV strain D1036/08. Phylogenetic analyses placed the two GoAdV strains monophyletically in the genus Aviadenovirus. We propose designating strains P29 and D1036/08 as GoAdV-4 and GoAdV-5, respectively.

Research paper thumbnail of Morphological and molecular biological studies on intramuscular Myxobolus spp. of cyprinid fish

Journal of Fish Diseases, 2002

The validity of Myxobolus species infecting the skeletal muscles of six cyprinid fish species was... more The validity of Myxobolus species infecting the skeletal muscles of six cyprinid fish species was studied by morphological and molecular biological methods. Intracellularly developing Myxobolus spores identified as M. cyprini from the common carp, M. musculi from the barbel, and M. pseudodispar from the roach, rudd, common bream and white bream were very similar in their shape and size. Nonetheless, in species identified as M. pseudodispar, the occurrence of spores with an asymmetrical shape was higher than in M. cyprini, while asymmetrical spores were only occasionally found in M. musculi. The DNA sequence analysis of the polymerase chain reaction (PCR)-amplified 18S rRNA gene of Myxobolus spores from these fish showed a similar phylogeny to that of their host species. As morphological studies and DNA sequence analysis demonstrated slight but real differences in the spores infecting muscles of the six cyprinid species, it is suggested that M. musculi, M. pseudodispar and M. cyprini are valid species.

Research paper thumbnail of Evidence of molecular evolution driven by recombination events influencing tropism in a novel human adenovirus that causes epidemic keratoconjunctivitis

PloS one, 2009

In 2005, a human adenovirus strain (formerly known as HAdV-D22/H8 but renamed here HAdV-D53) was ... more In 2005, a human adenovirus strain (formerly known as HAdV-D22/H8 but renamed here HAdV-D53) was isolated from an outbreak of epidemic keratoconjunctititis (EKC), a disease that is usually caused by HAdV-D8, -D19, or -D37, not HAdV-D22. To date, a complete change of tropism compared to the prototype has never been observed, although apparent recombinant strains of other viruses from species Human adenovirus D (HAdV-D) have been described. The complete genome of HAdV-D53 was sequenced to elucidate recombination events that lead to the emergence of a viable and highly virulent virus with a modified tropism. Bioinformatic and phylogenetic analyses of this genome demonstrate that this adenovirus is a recombinant of HAdV-D8 (including the fiber gene encoding the primary cellular receptor binding site), HAdV-D22, (the epsilon determinant of the hexon gene), HAdV-D37 (including the penton base gene encoding the secondary cellular receptor binding site), and at least one unknown or unsequen...

Research paper thumbnail of Molecular characterization of a lizard adenovirus reveals the first atadenovirus with two fiber genes and the first adenovirus with either one short or three long fibers per penton

Journal of virology, 2014

Although adenoviruses (AdVs) have been found in a wide variety of reptiles, including numerous sq... more Although adenoviruses (AdVs) have been found in a wide variety of reptiles, including numerous squamate species, turtles, and crocodiles, the number of reptilian adenovirus isolates is still scarce. The only fully sequenced reptilian adenovirus, snake adenovirus 1 (SnAdV-1), belongs to the Atadenovirus genus. Recently, two new atadenoviruses were isolated from a captive Gila monster (Heloderma suspectum) and Mexican beaded lizards (Heloderma horridum). Here we report the full genomic and proteomic characterization of the latter, designated lizard adenovirus 2 (LAdV-2). The double-stranded DNA (dsDNA) genome of LAdV-2 is 32,965 bp long, with an average G+C content of 44.16%. The overall arrangement and gene content of the LAdV-2 genome were largely concordant with those in other atadenoviruses, except for four novel open reading frames (ORFs) at the right end of the genome. Phylogeny reconstructions and plesiomorphic traits shared with SnAdV-1 further supported the assignment of LAdV...

Research paper thumbnail of DNA sequence of a small, unidentified plasmid isolated from a Haemophilus somnus strain: Short communication

Acta Veterinaria Hungarica

One of the plasmids present in a Haemophilus somnus strain isolated from nasal discharge of a cat... more One of the plasmids present in a Haemophilus somnus strain isolated from nasal discharge of a cattle with respiratory disease was purified and cloned for DNA sequencing. The plasmid was found to be 1065 base pairs long with 39.2% G + C content, and showed no homology to any DNA sequenced so far. It has no capacity to code any protein longer than 43 residues. It is not clear yet if this plasmid codes Haemophilus somnus specific factors.

Research paper thumbnail of Introduction of the family Alloherpesviridae: the first molecular detection of herpesviruses of cyprinid fish in Hungary

Magyar Allatorvosok Lapja

The taxonomy of herpesviruses has been changed recently radically. In 2008, a new order, the Herp... more The taxonomy of herpesviruses has been changed recently radically. In 2008, a new order, the Herpesvirales was established to contain the family Herpesviridae along with two additional new families, Alloherpesviridae and Malacoherpesviridae. The family Alloherpesviridae includes the herpesviruses isolated from amphibians and fish, whereas Malacoherpesviridae contains the single herpesvirus originating from an invertebrate host, namely an oyster. Previously, these viruses had been classified into the family Herpesviridae based on their characteristic morphology, although they did not fit unambiguously into any genus or subfamily of the family. The newly established family Alloherpesviridae is now officially divided into four genera. One of these contains the herpesviruses of amphibians, whereas the other three contain those of different fishes. The genus Cyprinivirus includes three virus species originating from cyprinid fishes. Cyprinid herpesvirus 1 (CyHV-1) is the causative agent ...

Research paper thumbnail of Crystal structure of the fibre head domain of bovine adenovirus 4, a ruminant atadenovirus

Virology journal, 2015

In adenoviruses, primary host cell recognition is generally performed by the head domains of thei... more In adenoviruses, primary host cell recognition is generally performed by the head domains of their homo-trimeric fibre proteins. This first interaction is reversible. A secondary, irreversible interaction subsequently takes place via other adenovirus capsid proteins and leads to a productive infection. Although many fibre head structures are known for human mastadenoviruses, not many animal adenovirus fibre head structures have been determined, especially not from those belonging to adenovirus genera other than Mastadenovirus. We constructed an expression vector for the fibre head domain from a ruminant atadenovirus, bovine adenovirus 4 (BAdV-4), consisting of amino acids 414-535, expressed the protein in Escherichia coli, purified it by metal affinity and cation exchange chromatography and crystallized it. The structure was solved using single isomorphous replacement plus anomalous dispersion of a mercury derivative and refined against native data that extended to 1.2 Å resolution....

Research paper thumbnail of Infectious canine hepatitis: detection of canine adenovirus type 1 by polymerase chain reaction

Acta veterinaria Hungarica, 1996

A primer pair and a polymerase chain reaction (PCR) method earlier tested for the detection of hu... more A primer pair and a polymerase chain reaction (PCR) method earlier tested for the detection of human and animal adenoviruses were used to demonstrate the presence of canine adenovirus type 1 (CAV-1) in tissue culture and clinical specimens. A simple procedure of sample preparation was elaborated making the PCR easily applicable in rapid confirmation of the diagnosis of infectious canine hepatitis.

Research paper thumbnail of Four new inverted terminal repeat sequences from bovine adenoviruses reveal striking differences in the length and content of the ITRs

Virus genes, 2001

The inverted terminal repeat (ITR) of the genome of four bovine adenovirus (BAdV) types have been... more The inverted terminal repeat (ITR) of the genome of four bovine adenovirus (BAdV) types have been sequenced, analysed and compared to the ITRs of other adenoviruses. The length of ITRs of the examined BAdVs ranged between 59 and 368 base pairs, thus the presently known longest adenovirus ITR sequence is from BAdV-10. The conserved motifs and characteristic sequence elements of the ITRs providing different binding sites for replicative proteins of viral and cellular origin seemed to be distributed according to the proposed genus classification of BAdVs. The ITRs of BAdV-10 share similarity with the members of the genus Mastadenovirus, while the ITRs of the other three sequenced serotypes (BAdV-4, 5 and strain Rus) which are candidate members of the genus Atadenovirus are very short and contain NFI and Sp1 binding sites only. The analysis of the new ITRs implied that the nucleotide sequence of the so-called core origin is highly preserved within the mastadenovirus genus only.

Research paper thumbnail of Restriction site mapping of bovine adenovirus type 1

Acta veterinaria Hungarica, 1990

Physical maps of the DNA of bovine adenovirus (BAV) type 1 were established using ApaI, BamHI, Bs... more Physical maps of the DNA of bovine adenovirus (BAV) type 1 were established using ApaI, BamHI, BstEII, EcoRI and KpnI enzymes. The size of the viral genome was found to be around 35,000 base pairs. The orientation of the maps was determined by hybridizing Southern blots of BAV-1 DNA with the cloned hexon gene region of BAV-3.

Research paper thumbnail of Siadenovirus

The Springer Index of Viruses, 2002

Research paper thumbnail of Sequence, Transcriptional Analysis, and Deletion of the Bovine Adenovirus Type 1 E3 Region

Virology, 1998

The early 3 (E3) transcriptional unit of human adenoviruses (HAV) encodes proteins that modulate ... more The early 3 (E3) transcriptional unit of human adenoviruses (HAV) encodes proteins that modulate host antiviral immune defenses. HAV E3 sequences are highly variable; different HAV groups encode phylogenetically unrelated proteins. The role of the E3 region of many human and animal adenoviruses is unknown because the sequences are unrelated to previously characterized viruses and the functions of proteins encoded by these regions have not been studied. We sequenced a portion of the bovine adenovirus serotype 1 (BAV-1) genome corresponding to the putative E3 region. This sequence was substantially different from other adenoviral E3 sequences, including those of two other bovine adenoviruses. However, two regions of putative sequence conservation were identified. BAV-1 E3 sequences were identified in early and late transcripts, but, unlike HAV, introns were not detected in the E3 region transcripts. Like HAV E3, a majority of the BAV-1 E3 region was not essential for growth in cell culture, as demonstrated by the construction of a recombinant BAV-1 lacking 60% of the putative E3 region.

Research paper thumbnail of Genomic characterization of human adenovirus 36, a putative obesity agent

Virus Research, 2010

Increased levels of serum antibody titers against human adenovirus 36 (HAdV-D36) are associated w... more Increased levels of serum antibody titers against human adenovirus 36 (HAdV-D36) are associated with human obesity and experimental obesity in laboratory animals. While HAdV-D36 has been studied as an infectious agent implicated in obesity for over a decade, the complete genome sequence and its analysis have yet to be reported. A detailed analysis of the genome sequence of HAdV-D36 may be important to understand its role in obesity. Genomic and bioinformatic comparisons with other HAdVs identified differences that suggested unique functions. Global pairwise genome alignment with all sequenced human adenovirus D (HAdV-D) genomes revealed areas of nonconserved sequences in the hexon, E3 CR1␤, E3 CR1␥, and fiber genes. Phylogenetic analysis of all HAdV-D36 proteins confirmed that this virus belongs to species Human adenovirus D. This genomic analysis of HAdV-D36 provides an important tool for comprehending the role that this unique adenovirus may play in human obesity. Low amino acid sequence identity in the E3 CR1␤ and CR1␥ genes may suggest distinctive roles for these proteins. Furthermore, the predicted molecular models of the HAdV-D36 fiber protein seem to implicate a unique tissue tropism for HAdV-D36.

Research paper thumbnail of Molecular characterization of a lizard adenovirus reveals the first atadenovirus with two fiber genes and the first adenovirus with either one short or three long fibers per penton

Journal of virology, 2014

Although adenoviruses (AdVs) have been found in a wide variety of reptiles, including numerous sq... more Although adenoviruses (AdVs) have been found in a wide variety of reptiles, including numerous squamate species, turtles, and crocodiles, the number of reptilian adenovirus isolates is still scarce. The only fully sequenced reptilian adenovirus, snake adenovirus 1 (SnAdV-1), belongs to the Atadenovirus genus. Recently, two new atadenoviruses were isolated from a captive Gila monster (Heloderma suspectum) and Mexican beaded lizards (Heloderma horridum). Here we report the full genomic and proteomic characterization of the latter, designated lizard adenovirus 2 (LAdV-2). The double-stranded DNA (dsDNA) genome of LAdV-2 is 32,965 bp long, with an average G+C content of 44.16%. The overall arrangement and gene content of the LAdV-2 genome were largely concordant with those in other atadenoviruses, except for four novel open reading frames (ORFs) at the right end of the genome. Phylogeny reconstructions and plesiomorphic traits shared with SnAdV-1 further supported the assignment of LAdV...

Research paper thumbnail of Sequencing and phylogenetic analysis of the protease gene, and genetic mapping of bovine adenovirus type 10 define its relatedness to other bovine adenoviruses

Virus Research, 1998

The complete genome of a bovine adenovirus (BAV) type 10 isolate was molecularly cloned and parti... more The complete genome of a bovine adenovirus (BAV) type 10 isolate was molecularly cloned and partially sequenced. The encoded proteins were predicted by computer analysis of the DNA sequences of the ends or the entire length of the cloned viral fragments, and thus a rough genetic map was constructed. The protease gene of BAV-10 was completely sequenced and used in phylogenetic analysis. Based on the results of the phylogenetic analysis, and the location and presence of certain genes thought to be specifically characteristic of subgroup 1 or subgroup 2 BAVs, it could be concluded that, in spite of the striking similarity in certain biological properties, BAV-10 is not related to subgroup 2 BAVs as originally described. It does not however fit clearly into subgroup 1 either, the members of which show closer relationship with human adenoviruses. BAV-10 therefore should best be considered as the first member of a third subgroup of BAVs.

Research paper thumbnail of A novel siadenovirus detected in the kidneys and liver of Gouldian finches (Erythura gouldiae)

Veterinary Microbiology, 2014

This article appeared in a journal published by Elsevier. The attached copy is furnished to the a... more This article appeared in a journal published by Elsevier. The attached copy is furnished to the author for internal non-commercial research and education use, including for instruction at the authors institution and sharing with colleagues.

Research paper thumbnail of Molecular typing of fowl adenoviruses, isolated in Hungary recently, reveals high diversity

Veterinary Microbiology, 2013

Molecular typing of 26 recent fowl adenovirus (FAdV) isolates obtained in Eastern Hungary between... more Molecular typing of 26 recent fowl adenovirus (FAdV) isolates obtained in Eastern Hungary between 2006 and 2011 was performed. The viruses were isolated on primary chicken embryo liver cell cultures from carcasses of chickens, sent for diagnostic investigation. The isolates were subjected to a PCR to amplify a fragment from the hexon gene. Sequence analysis of the amplicons revealed the presence of all the five FAdV species in the country, among them FAdV-B, which has been barely found in Hungary before. The three strains classified as FAdV-B might represent a novel type within this species, as their nucleotide sequence identity to strain 340 (type FAdV-5) is below the commonly accepted intratype limit. Most of the strains (63%) were classified into species FAdV-E (12 strains) and FAdV-D (7 strains) in the study. Four of the isolates proved to be the mixture of two adenovirus strains belonging to two different FAdV types, in three from these four cases, these two types even belonged to two different species. Analysing the pathological findings bolsters certain established connections between some FAdV types and disease conditions. Gizzard erosion was found in connection with FAdV-1 strains only, and inclusion body hepatitis with FAdV-D and -E strains.

Research paper thumbnail of Detection and Analysis of Six Lizard Adenoviruses by Consensus Primer PCR Provides Further Evidence of a Reptilian Origin for the Atadenoviruses

Journal of Virology, 2004

A consensus nested-PCR method was designed for investigation of the DNA polymerase gene of adenov... more A consensus nested-PCR method was designed for investigation of the DNA polymerase gene of adenoviruses. Gene fragments were amplified and sequenced from six novel adenoviruses from seven lizard species, including four species from which adenoviruses had not previously been reported. Host species included Gila monster, leopard gecko, fat-tail gecko, blue-tongued skink, Tokay gecko, bearded dragon, and mountain chameleon. This is the first sequence information from lizard adenoviruses. Phylogenetic analysis indicated that these viruses belong to the genus Atadenovirus, supporting the reptilian origin of atadenoviruses. This PCR method may be useful for obtaining templates for initial sequencing of novel adenoviruses.

Research paper thumbnail of Recognition and partial genome characterization by non-specific DNA amplification and PCR of a new siadenovirus species in a sample originating from Parus major, a great tit

Journal of Virological Methods, 2010

A seemingly novel siadenovirus species was detected by PCR and sequencing in the sample of a grea... more A seemingly novel siadenovirus species was detected by PCR and sequencing in the sample of a great tit (Parus major) found dead in Hungary. Since the genus Siadenovirus has very few known members so far, further study of the virus was intriguing not only from epizootiological but also from taxonomical aspects. The sample, which had been tested in another PCR survey previously, consisted of less than 50 l of extracted nucleic acid. To ensure sufficient target DNA for an extended study, the viral genome had to be preserved. To this end, the sample was subjected to a novel method of non-specific DNA amplification. Using the amplified DNA as target, different PCR and sequencing strategies were applied with consensus or specific primers for the study of the central genome part of the putative tit adenovirus. The sequence of supposedly one half (13,628 bp) of the genome was determined including eight full genes between the genes of the IVa2 and hexon proteins. The gene content of the viral genome fragment as well as the results of the phylogenetic analyses with different proteins confirmed the discovery of a new species in the genus Siadenovirus. This is the first report on the detection of an adenovirus in great tits. The methods, described in this work, proved suitable for the recovery of nucleic acid samples that contain irreplaceable microbial genomic DNA but are only available in limited quantities.

Research paper thumbnail of DNA sequencing and analysis of the right-hand part of the genome of the unique bovine adenovirus type 10

Journal of General Virology, 2004

DNA sequencing and analysis of the right-hand part of the genome of the unique bovine adenovirus ... more DNA sequencing and analysis of the right-hand part of the genome of the unique bovine adenovirus type 10

Research paper thumbnail of Genome sequence of a waterfowl aviadenovirus, goose adenovirus 4

Journal of General Virology, 2012

We present, to our knowledge, the first complete genome sequence of a waterfowl aviadenovirus, go... more We present, to our knowledge, the first complete genome sequence of a waterfowl aviadenovirus, goose adenovirus (GoAdV) strain P29, and an analysis of its genetic content in comparison with five published aviadenovirus genome sequences. Of the 35 genes predicted to encode functional proteins, the central region of the genome contains 19 (IVa2 to fiber-2) that were inherited from the ancestor of all known adenoviruses. Of the remaining genes, nine have orthologues only in aviadenoviruses and seven lack orthologues in any adenovirus. We also obtained limited sequence data for a pathogenic GoAdV strain D1036/08. Phylogenetic analyses placed the two GoAdV strains monophyletically in the genus Aviadenovirus. We propose designating strains P29 and D1036/08 as GoAdV-4 and GoAdV-5, respectively.

Research paper thumbnail of Morphological and molecular biological studies on intramuscular Myxobolus spp. of cyprinid fish

Journal of Fish Diseases, 2002

The validity of Myxobolus species infecting the skeletal muscles of six cyprinid fish species was... more The validity of Myxobolus species infecting the skeletal muscles of six cyprinid fish species was studied by morphological and molecular biological methods. Intracellularly developing Myxobolus spores identified as M. cyprini from the common carp, M. musculi from the barbel, and M. pseudodispar from the roach, rudd, common bream and white bream were very similar in their shape and size. Nonetheless, in species identified as M. pseudodispar, the occurrence of spores with an asymmetrical shape was higher than in M. cyprini, while asymmetrical spores were only occasionally found in M. musculi. The DNA sequence analysis of the polymerase chain reaction (PCR)-amplified 18S rRNA gene of Myxobolus spores from these fish showed a similar phylogeny to that of their host species. As morphological studies and DNA sequence analysis demonstrated slight but real differences in the spores infecting muscles of the six cyprinid species, it is suggested that M. musculi, M. pseudodispar and M. cyprini are valid species.

Research paper thumbnail of Evidence of molecular evolution driven by recombination events influencing tropism in a novel human adenovirus that causes epidemic keratoconjunctivitis

PloS one, 2009

In 2005, a human adenovirus strain (formerly known as HAdV-D22/H8 but renamed here HAdV-D53) was ... more In 2005, a human adenovirus strain (formerly known as HAdV-D22/H8 but renamed here HAdV-D53) was isolated from an outbreak of epidemic keratoconjunctititis (EKC), a disease that is usually caused by HAdV-D8, -D19, or -D37, not HAdV-D22. To date, a complete change of tropism compared to the prototype has never been observed, although apparent recombinant strains of other viruses from species Human adenovirus D (HAdV-D) have been described. The complete genome of HAdV-D53 was sequenced to elucidate recombination events that lead to the emergence of a viable and highly virulent virus with a modified tropism. Bioinformatic and phylogenetic analyses of this genome demonstrate that this adenovirus is a recombinant of HAdV-D8 (including the fiber gene encoding the primary cellular receptor binding site), HAdV-D22, (the epsilon determinant of the hexon gene), HAdV-D37 (including the penton base gene encoding the secondary cellular receptor binding site), and at least one unknown or unsequen...