A dicistronic gene pair within a cluster of “EF-hand” protein genes in the genomes of Drosophila species (original) (raw)
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Drosophila melanogaster contains a single calmodulin gene
Journal of Molecular Biology, 1990
We have previously characterized a calmodutin gene from the organism Drosophila melanogaster. In the higher vertebrates a multi-gene system for encoding calmodulin is present and, in at least one invertebrate species, genes encoding highly related calmodulin isotypes exist. We have therefore searched for additional calmodulin genes within D. melanogaster. Although our searches were sensitive enough to detect a relatively divergent gene encoding a calmodulin family protein, we were unable to detect any additional genes for calmodulin per se. Further studies of the structure and expression of the single calmodulin gene of D. melanogaster have established that the gene contains a tiny additional 5' exon encoding only 50 residues of the 5' leader. Sequencing at the 3' terminus has established that the two transcript size classes derived from the gene are produced as a result of alternative polyadenylation site usage. The relative abundance of the two size classes of mRNAs differs throughout the life cycle, indicating developmental regulation of polyadenylation site usage. A Single Calmodulin Gene in Drosophila melanogaster Most species possess a single protein sequence for the regulatory Ca2+-binding protein calmodulin (CAM §). It has been shown, however, that in vertebrates this single protein sequence is encoded by at least three different genes, a finding that is of unexplained physiological significance (Nojima &
Mechanisms of Development, 1994
Acp26Aa and Acp26Ab are Drosophila male accessory gland transcripts that are tightly linked and transcribed from the same DNA strand. Despite their being separated by 20 base pairs, the transcripts show identical responses to several developmental signals. These observations make it important to determine whether the 26A region contains two separable genes with the same developmental expression or a single developmentally regulated transcription unit whose product is processed to yield Acp26Aa and Acp26Ab. We show that Acp26Aa and Acp26Ab are separate mRNAs using a reverse transcription-polymerase chain reaction assay and reporter gene fusions. We also show that the regulatory elements for Acp26Ab lie within a fragment containing the intergenic region and transcribed sequences of Acp26Aa and Acp26Ab.
Origin and Evolution of a New Gene Expressed in the Drosophila Sperm Axoneme
Genetica, 2003
Sdic is a new gene that evolved recently in the lineage of Drosophila melanogaster. It was formed from a duplication and fusion of the gene AnnX, which encodes annexin X, and Cdic, which encodes the intermediate polypeptide chain of the cytoplasmic dynein. The fusion joins AnnX exon 4 with Cdic intron 3, which brings together three putative promoter elements for testes- specific expression of Sdic: the distal conserved element (DCE) and testes-specific element (TSE) are derived from AnnX, and the proximal conserved element (PCE) from Cdic intron 3. Sdic transcription initiates within the PCE, and translation is initiated within the sequence derived from Cdic intron 3, continuing through a 10 base pair insertion that creates a new splice donor site that enables the new coding sequence derived from intron 3 to be joined with the coding sequence of Cdic exon 4. A novel protein is created lacking 100 residues at the amino end that contain sequence motifs essential for the function of cytoplasmic dynein intermediate chains. Instead, the amino end is a hydrophobic region of 16 residues that resembles the amino end of axonemal dynein intermediate chains from other organisms. The downstream portion of Sdic features large deletions eliminating Cdic exons v2 and v3, as well as multiple frameshift deletions or insertions. The new protein becomes incorporated into the tail of the mature sperm and may function as an axonemal dynein intermediate chain. The new Sdic gene is present in about 10 tandem repeats between the wildtype Cdic and AnnX genes located near the base of the X chromosome. The implications of these findings are discussed relative to the origin of new gene functions and the process of speciation.
Mechanisms of Development, 1997
We identified and characterized the don juan gene (dj) of Drosophila melanogaster. The don juan gene codes for a sperm specific protein component with an unusual repetitive six amino acid motif (DPCKKK) in the carboxy-terminal part of the protein. The expression of Don Juan is limited to male germ cells where transcription of the dj gene is initiated during meiotic prophase. But Western blot experiments indicate that DJ protein occurs just postmeiotically. Examination of transgenic flies bearing a dj-promoter-lacZ reporter construct revealed lacZ mRNA distribution resembling the expression pattern of the endogenous dj mRNA in the adult testes, whereas /3-galactosidase expression is exclusively present in postmeiotic germ cells. Thus, these observations strongly suggest that dj transcripts are under translational repression until spermiogenesis. To study the function and subcellular distribution of DJ in spernliogenesis we expressed a chimaeric dj-GFP fusion gene in the male germline exhibiting strong GFP fluorescence in the live testes, where only elongated sperrnatids are decorated. With regard to the characteristic expression pattern of DJ protein and its conspicuous repeat units possible functional roles are discussed.
Analysis of Drosophila melanogaster testis transcriptome
BMC genomics, 2018
The formation of matured and individual sperm involves a series of molecular and spectacular morphological changes of the developing cysts in Drosophila melanogaster testis. Recent advances in RNA Sequencing (RNA-Seq) technology help us to understand the complexity of eukaryotic transcriptomes by dissecting different tissues and developmental stages of organisms. To gain a better understanding of cellular differentiation of spermatogenesis, we applied RNA-Seq to analyse the testis-specific transcriptome, including coding and non-coding genes. We isolated three different parts of the wild-type testis by dissecting and cutting the different regions: 1.) the apical region, which contains stem cells and developing spermatocytes 2.) the middle region, with enrichment of meiotic cysts 3.) the basal region, which contains elongated post-meiotic cysts with spermatids. Total RNA was isolated from each region and analysed by next-generation sequencing. We collected data from the annotated 174...
Cross-Species Comparison of Drosophila Male Accessory Gland Protein Genes
Genetics, 2005
Drosophila melanogaster males transfer seminal fluid proteins along with sperm during mating. Among these proteins, ACPs (Accessory gland proteins) from the male's accessory gland induce behavioral, physiological, and life span reduction in mated females and mediate sperm storage and utilization. A previous evolutionary EST screen in D. simulans identified partial cDNAs for 57 new candidate ACPs. Here we report the annotation and confirmation of the corresponding Acp genes in D. melanogaster. Of 57 new candidate Acp genes previously reported in D. melanogaster, 34 conform to our more stringent criteria for encoding putative male accessory gland extracellular proteins, thus bringing the total number of ACPs identified to 52 (34 plus 18 previously identified). This comprehensive set of Acp genes allows us to dissect the patterns of evolutionary change in a suite of proteins from a single male-specific reproductive tissue. We used sequence-based analysis to examine codon bias, gene...
Transcript maps of Drosophila yolk protein genes
Journal of Molecular Biology, 1982
We describe messenger RNA transcription maps of the three yolk protein (YP) genes of Drosophila melanogaster. Nuclease protection experiments demonstrated that the intron-exon structures of the YPl and YP2 genes are similar and have in the 5' to 3' direction: a 290 base-pair exon, a 65 to 80 base-pair intron and then a 1300 base-pair exon. The similarity between these two genes extends to their nucleotide sequence becaiise electron microscopy revealed that they form 930 basepair heteroduplexes. The YP3 gene is less similar. It has two 65 to 75 base-pair introns, which separate in the 5' to 3' direction exons of 240, 400 and 840 base-pairs. Electron microscopy revealed little homology between the YP3 and YP2 genes (two regions of approximately 70 and 150 base-pairs) and none between the YP3 and YPl genes. A single in vivo transcript was detected for each of the YPl and YP3 genes and two were detected for the YP2 gene. The YP2 transcripts, both present in polysomal RNA, differ in length by 80 base-pairs at their 3' ends. The YPl and YP2 genes are transcribed from opposite strands and their 5' ends are separated by 1'2 x 10^ base-pairs. Thus these two genes as well as the co-ordinately transcribed YP3 gene (Barnett & Wensink, 1981) have separate promoters. We conclude that when the steroid hormone, ecdysone, causes transcription of these genes to begin, it simultaneously influences three promoters.
A 14 bp promoter element directs the testis specificity of the Drosophila β2 tubulin gene
The EMBO Journal
To analyze the regulation of gene expression during male germ cell development, we investigated the testis-specific expression of the Drosophila (2 tubulin gene. Germ line transformation experiments with the upstream region of the D.melanogaster (32 tubulin gene fused to the Escherichia coli lacZ gene resulted in the correct tissue speciric expression of the reporter gene. Furthermore, we showed that the upstream sequences of the (2 tubulin gene of the distantly related species D.hydei can drive the expression of the lacZ gene testis specifically in D.melanogaster flies. A detailed deletion analysis showed that 53 bp of upstream and 23 bp (D.melanogaster) or 29 bp (D.hydei) of leader sequences are sufficient to confer tissue specificity. The short promoter regions contain a 14 bp motif at identical positions in both species, which acts as a position-dependent promoter element. In vitro mutagenesis and subsequent germline transformation experiments revealed that this sequence is the only element necessary for the testis-specific transcription of the (2 tubulin gene in Drosophila.
Genetics
A contiguous sequence of nearly 3 Mb from the genome of Drosophila melanogaster has been sequenced from a series of overlapping P1 and BAC clones. This region covers 69 chromosome polytene bands on chromosome arm 2L, including the genetically well-characterized "Adh region." A computational analysis of the sequence predicts 218 protein-coding genes, 11 tRNAs, and 17 transposable element sequences. At least 38 of the protein-coding genes are arranged in clusters of from 2 to 6 closely related genes, suggesting extensive tandem duplication. The gene density is one protein-coding gene every 13 kb; the transposable element density is one element every 171 kb. Of 73 genes in this region identified by genetic analysis, 49 have been located on the sequence; P-element insertions have been mapped to 43 genes. Ninety-five (44%) of the known and predicted genes match a Drosophila EST, and 144 (66%) have clear similarities to proteins in other organisms. Genes known to have mutant phe...
Male Accessory Gland Secretory Proteins in nasuta Subgroup of Drosophila: Synthetic Activity of Acp
Zoological Science, 2002
Male accessory gland secretory proteins in seven members of Drosophila ll aSlIla subgroup were analyzed by SDS -PAGE in combination with different staining techniques such as CBB-R2 50, Silver, PAS . PAS-s ilver and zinc-il11id azole reverse stai ni ng. Based on coom3ssie blue patterns the protein fractions cou ld be classified in to :1 maj or groups nal11cly group I, group II as well as gro up III ; with high molecu lar weigh t fractions falling into group I and low molecular weight fraction s into group III. All the three groups of fractions are post-translationally modified by way of glycosylatio n and group III frac ti ons are found to be highly glycosy lated. Fracti ons of groups I and II when locali zed with sil ve r slain and gruup III fractions when locali zed wi th PAS-si lver stai n appear ye ll ow; sugges tin g that th ey are sialoglycoprotcin s. A 40 kD fra ction of gro up II shows differential staining property wi th zinc-imidazo le stain in closely re latcd spec ies n:lI11ely D. II . 1111.1'11 111 :lI1 d D. II . alholllicalls. Analysis of thi s protein fraction in F1malcs of an int erspecifi c cross revealecl th at it is sylllhcs ized hy Xchromosomal gene.