Jebasingh Tennyson | Madurai Kamaraj University, Madurai, Tamil Nadu, INDIA (original) (raw)

Papers by Jebasingh Tennyson

Additional file 1: Figure S1. BBrMV miRNA2 sequencing chromatogram image. Table S1. Predicted BBr... more Additional file 1: Figure S1. BBrMV miRNA2 sequencing chromatogram image. Table S1. Predicted BBrMV miRNA targets and their functions. Figure S2. Gene Ontology (GO) analysis of the BBrMV miRNA2 target genes using Blast2GO. Figure S3. The multiple sequence alignment of the predicted BBrMV miRNA sequences among Philippines, Trichy, Wayanad and Ginger infecting BBrMV isolates. Table S2. Number of mismatches at mature miRNA sequences between the isolates of BBrMV. Figure S4. Stepwise virus miRNA prediction scheme. Figure S5. Schematic representation of stem-loop RT-PCR. Table S3. Primers to screen BBrMV miRNA sequences.

Archives of Virology, 2021

Nuclear inclusion a protease (NIaPro), a major protease of potyvirids, processes its cognate vira... more Nuclear inclusion a protease (NIaPro), a major protease of potyvirids, processes its cognate viral polyprotein at distinct cleavage sites. Although Potyviridae is the largest family of the realm Riboviria, the individual NIaPro enzymes and their cleavage sites are believed to be species-specific. In the present study, the NIaPro amino acid sequences of 165 potyvirids of 10 genera and their 1154 cleavage sites were compared to understand their genus/species-specificity and functional regulation. Of these, the NIaPro of macluraviruses, maintains a constant length of 217 amino acids, while those of other genera allow variation. In particular, poaceviruses exhibited a broad range of NIaPro amino acid sequence lengths. Alignment of 162 NIaPro amino acid sequences showed that the N- and C-terminal regions allow variations, while the central region, with the catalytic triad and S1 subsite, are highly conserved. NIaPro cleavage sites are composed of seven amino acids (heptapeptide) denoted as P6-P5-P4-P3-P2-P1/P1'. A survey of 1154 cleavage sites showed that the P1 position is predominantly occupied by Gln/Glu, as is seen in picornaviruses. The P6 (Glu), P4 (Val/Cys/Gln), P2 (His/Tyr/Leu), and P1' (Ser/Ala/Gly/Met) positions are predominantly occupied by genus-specific residues, while P5 and P3 are not genus-specific. The 6K2-VPg and VPg-NIaPro junctions possess Glu at the P1 position in order to maintain latency.

Potyviridae comprises more than 200 ssRNA viruses, many of which have a broad host range and geog... more Potyviridae comprises more than 200 ssRNA viruses, many of which have a broad host range and geographical distributions. Potyvirids (members of Potyviridae) infect several economically important plants such as saffron, cardamom, cucumber, pepper, potato, tomato, yam, etc. Cumulatively, potyvirids cause a substantial economic loss. The major bottleneck in developing an efficient antiviral strategy is that viruses quickly evade host immunity owing to their higher mutation and recombination rates. Due to this reason, the emergence of newer and improved broad-spectrum approaches to combat viral infections is essential. The use of microRNA's (miRNA) to circumvent viral infection against animal viruses has been successfully employed. Fewer studies reported the development of efficient miRNA-based antivirus resistant strategies against plant viruses and none focused on multiple virus resistance. We focused on potyviruses since studies are limited and identification of conserved miRNAs ...

ExRNA, 2020

Background Potyviridae is the largest plant infecting family under the monophyletic group Ribovir... more Background Potyviridae is the largest plant infecting family under the monophyletic group Riboviria, infects many of the food, fodder and ornamental crops. Due to the higher mutation and recombination rate, potyvirids are evolving rapidly, adapting to the environmental chaos and expanding their hosts. Virus control measures are need to be updated as the economic importance of potyvirids is massive. microRNAs (miRNAs) are well known for their functional importance in eukaryotes and many viruses. Regardless of its biogenesis, whether canonical or noncanonical, microRNA centric antivirus approaches attract the researchers to the hopeful future of next-generation broad-spectrum antiviral measures. Methods In this study, we predicted and screened banana bract mosaic virus (BBrMV) encoding miRNAs by computation approaches and their targets on banana transcriptome using plant small RNA target analysis server (psRNAtarget). The target gene functions were annotated by Blast2GO. The predicted...

Epigenomes, 2018

Bhendi yellow vein mosaic virus (BYVMV) belongs to the monopartite begomovirus associated with th... more Bhendi yellow vein mosaic virus (BYVMV) belongs to the monopartite begomovirus associated with the β satellite. As a single-stranded DNA (ssDNA) virus, it should be amenable to transcriptional and post-transcriptional gene silencing (TGS and PTGS). Previously, we had demonstrated C2, C4 and βC1 to be having different levels of influence on PTGS. Hence in the present study, a series of experiments such as agroinfiltration, chop-polymerase chain reaction (PCR), quantitative PCR (qPCR) and bisulfite next generation sequencing (NGS) were designed to analyse the involvement of BYVMV proteins on DNA methylation suppression. From the preliminary studies, we concluded that BYVMV genes were responsible for TGS suppression and C2, C4 genes from BYVMV were selected for further studies. Agroinfiltration experiments with mutant C2 and C4 partial tandem repeat (PTR) constructs of BYVMV have confirmed the role of C2 and C4 in DNA methylation impairment. The protoplast replication assay has shown that C4 was not an impediment for viral DNA replication and subsequent agroinfiltration studies with the C4 mutant BYVMV PTR construct have revealed the involvement of C4 in viral DNA movement.

Archives of Virology, 2018

We have analysed the genome sequence of Wuhan poty-like virus 1 (WuPLV1), reported as an unclassi... more We have analysed the genome sequence of Wuhan poty-like virus 1 (WuPLV1), reported as an unclassified RNA virus in GenBank (Accession no: KX884573.1). Based on the polyprotein sequence identity (ranging from 55.2 to 71.1%), with classifiable members of the Macluravirus genus of the plant virus family Potyviridae, we suggest that WuPLV1 represents a possible new species of Macluravirus, although the virus was isolated from the Chinese land snail Mastigeulota kiangsinensis, which is not known to be a host or vector of macluraviruses.

Archives of Virology, 2019

The complete genome sequence of the KS isolate of cardamom mosaic virus (CdMV) was determined usi... more The complete genome sequence of the KS isolate of cardamom mosaic virus (CdMV) was determined using transcriptome sequencing data from CdMV-infected Elettaria cardamomum as well as from overlapping cDNA clones made from RNA extracted from viral particles. The viral genome consists of 8249 nucleotides (nt) and encodes a large polyprotein of 2636 amino acids (aa). The polyprotein of CdMV shared 48.9%-67.4% aa sequence identity with other reported macluraviruses. Similar to the other members of genus Macluravirus, the genome of CdMV lacks the P1 coding region and the N-terminus of the HC-Pro coding region. The putative small open reading frame, PIPO, embedded within the P3 cistron, is preceded by a C(A) 6 motif instead of G(A) 6. Phylogenetic analysis based on the complete genome sequence aided the grouping of CdMV along with all other macluraviruses and showed that it is closely related to alpinia oxyphylla mosaic virus (AloMV). Among CdMV isolates, the KS isolate is most similar to the Appangala isolate based on disease symptoms and phylogeny.

Archives of Virology, 2018

Biocatalysis and Agricultural Biotechnology, 2018

Elettaria cardamomum is an economically important spice crop. Genomic analysis of cardamom is oft... more Elettaria cardamomum is an economically important spice crop. Genomic analysis of cardamom is often faced with the limitation of inefficient nucleic acid extraction due to its high content of polyphenols and polysaccharides. In this study, a highly efficient DNA and RNA extraction protocol for cryopreserved samples from small cardamom plant was developed with modification in the CTAB and SDS method for DNA and RNA, respectively, with the inclusion of 2% ascorbic acid. DNA isolated by this method is highly suitable for PCR, restriction digestion and RAPD analysis. The RNA extraction method described here represent the presence of plant mRNA, small RNAs and viral RNA and, the isolated RNA proved amenable for RT-PCR and amplification of small and viral RNA. Nucleic acids extraction protocol developed here will be useful to develop genetic marker for cardamom, to clone cardamom genes, small RNAs and cardamom infecting viral genes and to perform gene expression and small RNA analysis.

FEMS microbiology letters, Jan 11, 2018

sRNAs are a class of gene regulators in bacteria, playing a central role in its response to envir... more sRNAs are a class of gene regulators in bacteria, playing a central role in its response to environmental changes. Bioinformatic prediction facilitates the identification of sRNAs expressed at different conditions. We propose a novel method of prediction of sRNAs from the genome of Agrobacterium based on PWM matrix of conditional sigma factors. sRNAs predicted from the genome are integrated with the virulence specific transcriptome data to identify putative sRNAs that are over expressed during Agrobacterial virulence induction. A total of 384 sRNAs are predicted from transcriptome data analysis of Agrobacterium fabrum and 100-500 sRNAs from the genome of different Agrobacterial strains. In order to refine our study, a final set of 10 novel sRNAs with best features across different replicons targeting virulence genes are experimentally identified using semi-quantitative PCR. Since Ti plasmid plays major role in virulence, out of 10 sRNAs across the replicons, 4 novel sRNAs differenti...

Biochimica et biophysica acta, Jan 12, 2018

Leptospira, the causative agent of leptospirosis is known to have many proteases with potential t... more Leptospira, the causative agent of leptospirosis is known to have many proteases with potential to degrade extracellular matrix. However, a multipronged approach to identify, classify, characterize and elucidate their role has not been attempted. Our proteomic approach using high-resolution LC-MS/MS analysis of Triton X-114 fractions of Leptospira interrogans resulted in the identification of 104 proteases out of 130 proteases predicted by MEROPS. In Leptospira approximately 3.5% of the genome complements for proteases, which include catalytic types of metallo-, serine-, cysteine-, aspartic-, threonine- and asparagine- peptidases. Comparison of proteases from different serovars revealed that M04, M09B, M14A, M75, M28A, A01 and U73 protease families are exclusively present in pathogenic form. The M23 and S33 protease families are represented with >14 members in Leptospira. The differential expression under physiological temperature (37 °C) and osmolarity (300 mOsM) showed that pro...

Biochemical Genetics, 2022

The involvement of many putative genetic factors makes osteoporosis a complex disease. With incre... more The involvement of many putative genetic factors makes osteoporosis a complex disease. With increasing longevity of the Indian population, it's now being realized that, as within the West, osteoporotic fractures are also a significant explanation for morbidity and mortality in postmenopausal women. Studies have suggested that the genetic component liable for bone mass could be linked to single nucleotide polymorphisms. Therefore, this study is aimed to research the role of seven gene polymorphisms previously associated with bone phenotype in a cohort of postmenopausal South Indian women from Tamil Nadu. The subjects for the study (n = 300) included 100 osteoporotic women (age 59.3 ± 9.26), 100 osteopenic women (age 55.6 ± 8.17) and 100 non-osteoporotic women as controls (age 55.4 ± 8.85).Genetic polymorphisms were determined by polymerase chain reaction (PCR)-restriction fragment length polymorphism. Case-control genetic association analysis of BsmI of the VDR and BstBI of the PTH gene showed a significant allelic association with low bone mineral density amongst the osteoporotic postmenopausal women. The association of BMD with the VDR gene polymorphisms revealed that the average BMD in the BsmI polymorphism with the recessive genotype GG in osteoporotic women was significantly reduced compared with the average BMD in osteoporotic women with AA and AG genotypes. In the BstBI polymorphism, the BMD in the osteoporotic subjects were significantly lower in the AA group than in the GA and GG groups. These results provide evidence for an independent association between BMD and rs1544410 in VDR and rs6254 in PTH and may contribute in being a possible genetic marker for predicting the disease susceptibility in the population tested.

Additional file 1: Supplementary file 1 List of Accession numbers of Rhizobium etli with their re... more Additional file 1: Supplementary file 1 List of Accession numbers of Rhizobium etli with their respective replicons used in our study. Supplementary file 2 Position Weight Matrix (PWM) log-odds ratio of nucleotides at each position of the sigma factor 32 binding motif. Supplementary file 3 Consensus sequence logos of sigma 32 matrix used for sRNA Scanner program. Supplementary file 4 List of sRNAs overlapped with the published data. a) Overlapped sequences with Vercruysse et al. (2010) data, b) with López-Leal et al. (2014) data. Supplementary file 5 List of flanking genes of the predicted common sRNAs under heat shock and saline shock conditions and sigma factor 32 based of Rhizobium etli. Supplementary file 6 Promoter, terminator and secondary structures of the predicted common sRNAs from the genome and transcriptome data.

PROTEOMICS, 2020

The Triton X‐114‐based solubilization and temperature‐dependent phase separation of proteins is u... more The Triton X‐114‐based solubilization and temperature‐dependent phase separation of proteins is used for subcellular fractionation where, aqueous, detergent, and pellet fractions represents cytoplasmic, outer membrane (OM), and inner membrane proteins, respectively. Mass spectrometry‐based proteomic analysis of Triton X‐114 fractions of proteomic analysis of Leptospira interrogans identified 2957 unique proteins distributed across the fractions. The results are compared with bioinformatics predictions on their subcellular localization and pathogenic nature. Analysis of the distribution of proteins across the Triton X‐114 fractions with the predicted characteristics is performed based on “number” of unique type of proteins, and “quantity” which represents the amount of unique protein. The highest number of predicted outer membrane proteins (OMPs) and pathogenic proteins are found in aqueous and pellet fractions, whereas detergent fraction representing the OM has the highest quantity of OMPs and pathogenic proteins though lower in number than the aqueous and pellet fractions. This leaves the possibility of an upsurge in pathogenic proteins and OMPs on the OM under pathogenic conditions suggesting their potential use to combat leptospirosis. Further, the Triton X‐114 subcellular fractions are more correlated to enrichment of pathogenic proteins predicted by MP3 software than predicted localization.

Rhizobium-legume symbiosis is considered as the major contributor of biological nitrogen fixation... more Rhizobium-legume symbiosis is considered as the major contributor of biological nitrogen fixation. In the present study, we have identified sigma factor 54-regulated sRNAs from the genome of five Rhizobium strains and integrated with the free-living and symbiotic specific transcriptome data to identify the novel putative sRNAs that are over expressed during the regulation of nitrogen fixation. A total of 1059 sRNAs were predicted from each genome of the select set of Rhizobium strains and 1,375 sRNAs were predicted from the transcriptome data of Bradyrhizobium japonicum. Target mRNA analysis revealed the functional role of putative novel sRNAs from different free-living and symbiotic strains. Those novel sRNAs were inferred to target several nodulation and nitrogen fixation genes including nodC, nodJ, nodY, nodJ, nodM, nodW, nodZ, nifD, nifN, nifQ, fixK, fixL, Fdx, nolB, and several cytochrome proteins. Further, sRNAs of Bradyrhizobium japonicum which targeted the regulatory genes o...

Loa22 is OmpA-like outer membrane protein from Leptospira interrogans characterized in the C-term... more Loa22 is OmpA-like outer membrane protein from Leptospira interrogans characterized in the C-terminus domain which play an important role in the infection and immunological responses of leptospirosis. [1,2]. Phylogenetic tree was constructed for Loa22 by comparing it with Lipoproteins (LipL 71, 45, 41, 31, 32, 21 and 48) and with outer membrane proteins (OmpL 1, 47, 54, 36, 37). The comprising lineages of Loa 22 have largely varying rates of evolution. In the present study the physicochemical properties of Loa22 were assessed. The structure of Loa22 was predicted by threading method in RaptorX server and the structure was energy minimized and validated by using SAVES server. The stereochemical quality and the protein backbone conformations were done by Ramachandran Plot analysis. Four sequential and conformational B cell epitopes of Loa22 were found in Ellipro server and mapped to find the sequential epitopes. The same B cell linear epitopes was also predicted by ABC Pred, BepiPred ...

ExRNA, 2020

Background: Small non-coding RNAs (sRNAs) are regulatory molecules, present in all forms of life,... more Background: Small non-coding RNAs (sRNAs) are regulatory molecules, present in all forms of life, known to regulate various biological processes in response to the different environmental signals. In recent years, deep sequencing and various other computational prediction methods have been employed to identify and analyze sRNAs. Results: In the present study, we have applied an improved sRNA scanner method to predict sRNAs from the genome of Rhizobium etli, based on PWM matrix of conditional sigma factor 32. sRNAs predicted from the genome are integrated with the available stress specific transcriptome data to predict putative conditional specific sRNAs. A total of 271 sRNAs from the genome and 173 sRNAs from the transcriptome are computationally predicted. Of these, 25 sRNAs are found in both genome and transcriptome data. Putative targets for these sRNAs are predicted using TargetRNA2 and these targets are involved in a wide array of cellular functions such as cell division, transport and metabolism of amino acids, carbohydrates, energy production and conversion, translation, cell wall/membrane biogenesis, posttranslation modification, protein turnover and chaperones. Predicted targets are functionally classified based on COG analysis and GO annotations. Conclusion: sRNAs predicted from the genome, using PWM matrices for conditional sigma factor 32 could be a better method to identify the conditional specific sRNAs which expand the list of putative sRNAs from the intergenic regions (IgRs) of R. etli and closely related α-proteobacteria. sRNAs identified in this study would be helpful to explore their regulatory role in biological cellular process during the stress.

3 Biotech, 2019

Coccinia mosaic Virdhunagar virus (KY860899), Tomato leaf curl New Delhi virus (KY860898) and Tom... more Coccinia mosaic Virdhunagar virus (KY860899), Tomato leaf curl New Delhi virus (KY860898) and Tomato leaf curl Virdhunagar alphasatellite (KY848691) were found to be associated with leaf curl disease in Momordica charantia (bitter gourd). The complete nucleotide sequence of Coccinia mosaic Virdhunagar virus showed 82% identity with Coccinia mosaic Tamil Nadu virus (KM244719), whereas Tomato leaf curl New Delhi virus was 96% identical to Tomato leaf curl New Delhi virus (KP868764) and Tomato leaf curl Virdhunagar alphasatellite illustrated 81% similarity with Tomato leaf curl New Delhi alphasatellite (JQ041697). Phylogenetic and RDP analysis revealed the proximity of these begomoviruses with other monopartite begomoviruses and alphasatellites already reported from India. As per the threshold criteria laid down by International Committee on Taxonomy of Viruses for species demarcation in begomoviruses and satellite molecules, the identified virus isolates, Coccinia mosaic Virdhunagar virus and Tomato leaf curl Virdhunagar alphasatellite are proposed as new species. To the best of our knowledge, this is the first ever account of mixed infection of begomoviruses in Momordica charantia, a vegetable crop commonly cultivated throughout India.

Additional file 1: Figure S1. BBrMV miRNA2 sequencing chromatogram image. Table S1. Predicted BBr... more Additional file 1: Figure S1. BBrMV miRNA2 sequencing chromatogram image. Table S1. Predicted BBrMV miRNA targets and their functions. Figure S2. Gene Ontology (GO) analysis of the BBrMV miRNA2 target genes using Blast2GO. Figure S3. The multiple sequence alignment of the predicted BBrMV miRNA sequences among Philippines, Trichy, Wayanad and Ginger infecting BBrMV isolates. Table S2. Number of mismatches at mature miRNA sequences between the isolates of BBrMV. Figure S4. Stepwise virus miRNA prediction scheme. Figure S5. Schematic representation of stem-loop RT-PCR. Table S3. Primers to screen BBrMV miRNA sequences.

Archives of Virology, 2021

Nuclear inclusion a protease (NIaPro), a major protease of potyvirids, processes its cognate vira... more Nuclear inclusion a protease (NIaPro), a major protease of potyvirids, processes its cognate viral polyprotein at distinct cleavage sites. Although Potyviridae is the largest family of the realm Riboviria, the individual NIaPro enzymes and their cleavage sites are believed to be species-specific. In the present study, the NIaPro amino acid sequences of 165 potyvirids of 10 genera and their 1154 cleavage sites were compared to understand their genus/species-specificity and functional regulation. Of these, the NIaPro of macluraviruses, maintains a constant length of 217 amino acids, while those of other genera allow variation. In particular, poaceviruses exhibited a broad range of NIaPro amino acid sequence lengths. Alignment of 162 NIaPro amino acid sequences showed that the N- and C-terminal regions allow variations, while the central region, with the catalytic triad and S1 subsite, are highly conserved. NIaPro cleavage sites are composed of seven amino acids (heptapeptide) denoted as P6-P5-P4-P3-P2-P1/P1'. A survey of 1154 cleavage sites showed that the P1 position is predominantly occupied by Gln/Glu, as is seen in picornaviruses. The P6 (Glu), P4 (Val/Cys/Gln), P2 (His/Tyr/Leu), and P1' (Ser/Ala/Gly/Met) positions are predominantly occupied by genus-specific residues, while P5 and P3 are not genus-specific. The 6K2-VPg and VPg-NIaPro junctions possess Glu at the P1 position in order to maintain latency.

Potyviridae comprises more than 200 ssRNA viruses, many of which have a broad host range and geog... more Potyviridae comprises more than 200 ssRNA viruses, many of which have a broad host range and geographical distributions. Potyvirids (members of Potyviridae) infect several economically important plants such as saffron, cardamom, cucumber, pepper, potato, tomato, yam, etc. Cumulatively, potyvirids cause a substantial economic loss. The major bottleneck in developing an efficient antiviral strategy is that viruses quickly evade host immunity owing to their higher mutation and recombination rates. Due to this reason, the emergence of newer and improved broad-spectrum approaches to combat viral infections is essential. The use of microRNA's (miRNA) to circumvent viral infection against animal viruses has been successfully employed. Fewer studies reported the development of efficient miRNA-based antivirus resistant strategies against plant viruses and none focused on multiple virus resistance. We focused on potyviruses since studies are limited and identification of conserved miRNAs ...

ExRNA, 2020

Background Potyviridae is the largest plant infecting family under the monophyletic group Ribovir... more Background Potyviridae is the largest plant infecting family under the monophyletic group Riboviria, infects many of the food, fodder and ornamental crops. Due to the higher mutation and recombination rate, potyvirids are evolving rapidly, adapting to the environmental chaos and expanding their hosts. Virus control measures are need to be updated as the economic importance of potyvirids is massive. microRNAs (miRNAs) are well known for their functional importance in eukaryotes and many viruses. Regardless of its biogenesis, whether canonical or noncanonical, microRNA centric antivirus approaches attract the researchers to the hopeful future of next-generation broad-spectrum antiviral measures. Methods In this study, we predicted and screened banana bract mosaic virus (BBrMV) encoding miRNAs by computation approaches and their targets on banana transcriptome using plant small RNA target analysis server (psRNAtarget). The target gene functions were annotated by Blast2GO. The predicted...

Epigenomes, 2018

Bhendi yellow vein mosaic virus (BYVMV) belongs to the monopartite begomovirus associated with th... more Bhendi yellow vein mosaic virus (BYVMV) belongs to the monopartite begomovirus associated with the β satellite. As a single-stranded DNA (ssDNA) virus, it should be amenable to transcriptional and post-transcriptional gene silencing (TGS and PTGS). Previously, we had demonstrated C2, C4 and βC1 to be having different levels of influence on PTGS. Hence in the present study, a series of experiments such as agroinfiltration, chop-polymerase chain reaction (PCR), quantitative PCR (qPCR) and bisulfite next generation sequencing (NGS) were designed to analyse the involvement of BYVMV proteins on DNA methylation suppression. From the preliminary studies, we concluded that BYVMV genes were responsible for TGS suppression and C2, C4 genes from BYVMV were selected for further studies. Agroinfiltration experiments with mutant C2 and C4 partial tandem repeat (PTR) constructs of BYVMV have confirmed the role of C2 and C4 in DNA methylation impairment. The protoplast replication assay has shown that C4 was not an impediment for viral DNA replication and subsequent agroinfiltration studies with the C4 mutant BYVMV PTR construct have revealed the involvement of C4 in viral DNA movement.

Archives of Virology, 2018

We have analysed the genome sequence of Wuhan poty-like virus 1 (WuPLV1), reported as an unclassi... more We have analysed the genome sequence of Wuhan poty-like virus 1 (WuPLV1), reported as an unclassified RNA virus in GenBank (Accession no: KX884573.1). Based on the polyprotein sequence identity (ranging from 55.2 to 71.1%), with classifiable members of the Macluravirus genus of the plant virus family Potyviridae, we suggest that WuPLV1 represents a possible new species of Macluravirus, although the virus was isolated from the Chinese land snail Mastigeulota kiangsinensis, which is not known to be a host or vector of macluraviruses.

Archives of Virology, 2019

The complete genome sequence of the KS isolate of cardamom mosaic virus (CdMV) was determined usi... more The complete genome sequence of the KS isolate of cardamom mosaic virus (CdMV) was determined using transcriptome sequencing data from CdMV-infected Elettaria cardamomum as well as from overlapping cDNA clones made from RNA extracted from viral particles. The viral genome consists of 8249 nucleotides (nt) and encodes a large polyprotein of 2636 amino acids (aa). The polyprotein of CdMV shared 48.9%-67.4% aa sequence identity with other reported macluraviruses. Similar to the other members of genus Macluravirus, the genome of CdMV lacks the P1 coding region and the N-terminus of the HC-Pro coding region. The putative small open reading frame, PIPO, embedded within the P3 cistron, is preceded by a C(A) 6 motif instead of G(A) 6. Phylogenetic analysis based on the complete genome sequence aided the grouping of CdMV along with all other macluraviruses and showed that it is closely related to alpinia oxyphylla mosaic virus (AloMV). Among CdMV isolates, the KS isolate is most similar to the Appangala isolate based on disease symptoms and phylogeny.

Archives of Virology, 2018

Biocatalysis and Agricultural Biotechnology, 2018

Elettaria cardamomum is an economically important spice crop. Genomic analysis of cardamom is oft... more Elettaria cardamomum is an economically important spice crop. Genomic analysis of cardamom is often faced with the limitation of inefficient nucleic acid extraction due to its high content of polyphenols and polysaccharides. In this study, a highly efficient DNA and RNA extraction protocol for cryopreserved samples from small cardamom plant was developed with modification in the CTAB and SDS method for DNA and RNA, respectively, with the inclusion of 2% ascorbic acid. DNA isolated by this method is highly suitable for PCR, restriction digestion and RAPD analysis. The RNA extraction method described here represent the presence of plant mRNA, small RNAs and viral RNA and, the isolated RNA proved amenable for RT-PCR and amplification of small and viral RNA. Nucleic acids extraction protocol developed here will be useful to develop genetic marker for cardamom, to clone cardamom genes, small RNAs and cardamom infecting viral genes and to perform gene expression and small RNA analysis.

FEMS microbiology letters, Jan 11, 2018

sRNAs are a class of gene regulators in bacteria, playing a central role in its response to envir... more sRNAs are a class of gene regulators in bacteria, playing a central role in its response to environmental changes. Bioinformatic prediction facilitates the identification of sRNAs expressed at different conditions. We propose a novel method of prediction of sRNAs from the genome of Agrobacterium based on PWM matrix of conditional sigma factors. sRNAs predicted from the genome are integrated with the virulence specific transcriptome data to identify putative sRNAs that are over expressed during Agrobacterial virulence induction. A total of 384 sRNAs are predicted from transcriptome data analysis of Agrobacterium fabrum and 100-500 sRNAs from the genome of different Agrobacterial strains. In order to refine our study, a final set of 10 novel sRNAs with best features across different replicons targeting virulence genes are experimentally identified using semi-quantitative PCR. Since Ti plasmid plays major role in virulence, out of 10 sRNAs across the replicons, 4 novel sRNAs differenti...

Biochimica et biophysica acta, Jan 12, 2018

Leptospira, the causative agent of leptospirosis is known to have many proteases with potential t... more Leptospira, the causative agent of leptospirosis is known to have many proteases with potential to degrade extracellular matrix. However, a multipronged approach to identify, classify, characterize and elucidate their role has not been attempted. Our proteomic approach using high-resolution LC-MS/MS analysis of Triton X-114 fractions of Leptospira interrogans resulted in the identification of 104 proteases out of 130 proteases predicted by MEROPS. In Leptospira approximately 3.5% of the genome complements for proteases, which include catalytic types of metallo-, serine-, cysteine-, aspartic-, threonine- and asparagine- peptidases. Comparison of proteases from different serovars revealed that M04, M09B, M14A, M75, M28A, A01 and U73 protease families are exclusively present in pathogenic form. The M23 and S33 protease families are represented with >14 members in Leptospira. The differential expression under physiological temperature (37 °C) and osmolarity (300 mOsM) showed that pro...

Biochemical Genetics, 2022

The involvement of many putative genetic factors makes osteoporosis a complex disease. With incre... more The involvement of many putative genetic factors makes osteoporosis a complex disease. With increasing longevity of the Indian population, it's now being realized that, as within the West, osteoporotic fractures are also a significant explanation for morbidity and mortality in postmenopausal women. Studies have suggested that the genetic component liable for bone mass could be linked to single nucleotide polymorphisms. Therefore, this study is aimed to research the role of seven gene polymorphisms previously associated with bone phenotype in a cohort of postmenopausal South Indian women from Tamil Nadu. The subjects for the study (n = 300) included 100 osteoporotic women (age 59.3 ± 9.26), 100 osteopenic women (age 55.6 ± 8.17) and 100 non-osteoporotic women as controls (age 55.4 ± 8.85).Genetic polymorphisms were determined by polymerase chain reaction (PCR)-restriction fragment length polymorphism. Case-control genetic association analysis of BsmI of the VDR and BstBI of the PTH gene showed a significant allelic association with low bone mineral density amongst the osteoporotic postmenopausal women. The association of BMD with the VDR gene polymorphisms revealed that the average BMD in the BsmI polymorphism with the recessive genotype GG in osteoporotic women was significantly reduced compared with the average BMD in osteoporotic women with AA and AG genotypes. In the BstBI polymorphism, the BMD in the osteoporotic subjects were significantly lower in the AA group than in the GA and GG groups. These results provide evidence for an independent association between BMD and rs1544410 in VDR and rs6254 in PTH and may contribute in being a possible genetic marker for predicting the disease susceptibility in the population tested.

Additional file 1: Supplementary file 1 List of Accession numbers of Rhizobium etli with their re... more Additional file 1: Supplementary file 1 List of Accession numbers of Rhizobium etli with their respective replicons used in our study. Supplementary file 2 Position Weight Matrix (PWM) log-odds ratio of nucleotides at each position of the sigma factor 32 binding motif. Supplementary file 3 Consensus sequence logos of sigma 32 matrix used for sRNA Scanner program. Supplementary file 4 List of sRNAs overlapped with the published data. a) Overlapped sequences with Vercruysse et al. (2010) data, b) with López-Leal et al. (2014) data. Supplementary file 5 List of flanking genes of the predicted common sRNAs under heat shock and saline shock conditions and sigma factor 32 based of Rhizobium etli. Supplementary file 6 Promoter, terminator and secondary structures of the predicted common sRNAs from the genome and transcriptome data.

PROTEOMICS, 2020

The Triton X‐114‐based solubilization and temperature‐dependent phase separation of proteins is u... more The Triton X‐114‐based solubilization and temperature‐dependent phase separation of proteins is used for subcellular fractionation where, aqueous, detergent, and pellet fractions represents cytoplasmic, outer membrane (OM), and inner membrane proteins, respectively. Mass spectrometry‐based proteomic analysis of Triton X‐114 fractions of proteomic analysis of Leptospira interrogans identified 2957 unique proteins distributed across the fractions. The results are compared with bioinformatics predictions on their subcellular localization and pathogenic nature. Analysis of the distribution of proteins across the Triton X‐114 fractions with the predicted characteristics is performed based on “number” of unique type of proteins, and “quantity” which represents the amount of unique protein. The highest number of predicted outer membrane proteins (OMPs) and pathogenic proteins are found in aqueous and pellet fractions, whereas detergent fraction representing the OM has the highest quantity of OMPs and pathogenic proteins though lower in number than the aqueous and pellet fractions. This leaves the possibility of an upsurge in pathogenic proteins and OMPs on the OM under pathogenic conditions suggesting their potential use to combat leptospirosis. Further, the Triton X‐114 subcellular fractions are more correlated to enrichment of pathogenic proteins predicted by MP3 software than predicted localization.

Rhizobium-legume symbiosis is considered as the major contributor of biological nitrogen fixation... more Rhizobium-legume symbiosis is considered as the major contributor of biological nitrogen fixation. In the present study, we have identified sigma factor 54-regulated sRNAs from the genome of five Rhizobium strains and integrated with the free-living and symbiotic specific transcriptome data to identify the novel putative sRNAs that are over expressed during the regulation of nitrogen fixation. A total of 1059 sRNAs were predicted from each genome of the select set of Rhizobium strains and 1,375 sRNAs were predicted from the transcriptome data of Bradyrhizobium japonicum. Target mRNA analysis revealed the functional role of putative novel sRNAs from different free-living and symbiotic strains. Those novel sRNAs were inferred to target several nodulation and nitrogen fixation genes including nodC, nodJ, nodY, nodJ, nodM, nodW, nodZ, nifD, nifN, nifQ, fixK, fixL, Fdx, nolB, and several cytochrome proteins. Further, sRNAs of Bradyrhizobium japonicum which targeted the regulatory genes o...

Loa22 is OmpA-like outer membrane protein from Leptospira interrogans characterized in the C-term... more Loa22 is OmpA-like outer membrane protein from Leptospira interrogans characterized in the C-terminus domain which play an important role in the infection and immunological responses of leptospirosis. [1,2]. Phylogenetic tree was constructed for Loa22 by comparing it with Lipoproteins (LipL 71, 45, 41, 31, 32, 21 and 48) and with outer membrane proteins (OmpL 1, 47, 54, 36, 37). The comprising lineages of Loa 22 have largely varying rates of evolution. In the present study the physicochemical properties of Loa22 were assessed. The structure of Loa22 was predicted by threading method in RaptorX server and the structure was energy minimized and validated by using SAVES server. The stereochemical quality and the protein backbone conformations were done by Ramachandran Plot analysis. Four sequential and conformational B cell epitopes of Loa22 were found in Ellipro server and mapped to find the sequential epitopes. The same B cell linear epitopes was also predicted by ABC Pred, BepiPred ...

ExRNA, 2020

Background: Small non-coding RNAs (sRNAs) are regulatory molecules, present in all forms of life,... more Background: Small non-coding RNAs (sRNAs) are regulatory molecules, present in all forms of life, known to regulate various biological processes in response to the different environmental signals. In recent years, deep sequencing and various other computational prediction methods have been employed to identify and analyze sRNAs. Results: In the present study, we have applied an improved sRNA scanner method to predict sRNAs from the genome of Rhizobium etli, based on PWM matrix of conditional sigma factor 32. sRNAs predicted from the genome are integrated with the available stress specific transcriptome data to predict putative conditional specific sRNAs. A total of 271 sRNAs from the genome and 173 sRNAs from the transcriptome are computationally predicted. Of these, 25 sRNAs are found in both genome and transcriptome data. Putative targets for these sRNAs are predicted using TargetRNA2 and these targets are involved in a wide array of cellular functions such as cell division, transport and metabolism of amino acids, carbohydrates, energy production and conversion, translation, cell wall/membrane biogenesis, posttranslation modification, protein turnover and chaperones. Predicted targets are functionally classified based on COG analysis and GO annotations. Conclusion: sRNAs predicted from the genome, using PWM matrices for conditional sigma factor 32 could be a better method to identify the conditional specific sRNAs which expand the list of putative sRNAs from the intergenic regions (IgRs) of R. etli and closely related α-proteobacteria. sRNAs identified in this study would be helpful to explore their regulatory role in biological cellular process during the stress.

3 Biotech, 2019

Coccinia mosaic Virdhunagar virus (KY860899), Tomato leaf curl New Delhi virus (KY860898) and Tom... more Coccinia mosaic Virdhunagar virus (KY860899), Tomato leaf curl New Delhi virus (KY860898) and Tomato leaf curl Virdhunagar alphasatellite (KY848691) were found to be associated with leaf curl disease in Momordica charantia (bitter gourd). The complete nucleotide sequence of Coccinia mosaic Virdhunagar virus showed 82% identity with Coccinia mosaic Tamil Nadu virus (KM244719), whereas Tomato leaf curl New Delhi virus was 96% identical to Tomato leaf curl New Delhi virus (KP868764) and Tomato leaf curl Virdhunagar alphasatellite illustrated 81% similarity with Tomato leaf curl New Delhi alphasatellite (JQ041697). Phylogenetic and RDP analysis revealed the proximity of these begomoviruses with other monopartite begomoviruses and alphasatellites already reported from India. As per the threshold criteria laid down by International Committee on Taxonomy of Viruses for species demarcation in begomoviruses and satellite molecules, the identified virus isolates, Coccinia mosaic Virdhunagar virus and Tomato leaf curl Virdhunagar alphasatellite are proposed as new species. To the best of our knowledge, this is the first ever account of mixed infection of begomoviruses in Momordica charantia, a vegetable crop commonly cultivated throughout India.