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Comparative studies of allozyme loci in Drosophila simulans and Drosophila melanogaster. I. Three dipeptidase loci.

Genetic variation at three dipeptidase loci (Dip-A, Dip-B, and Dip-C) in Drosophila simulans was analyzed by starch gel electrophoresis. Dip-A was found to be polymorphic in four populations, while Dip-B and Dip-C were found to be polymorphic in one. The numbers of different alleles found at each respective locus were: Dip-A, two; Dip-B, two; and Dip-C, three. Dip-A was genetically mapped at 57.9 on the second chromosome, and Dip-B and Dip-C at 80.9 and 87.9 on the third chromosome, respectively. Neither Dip-B nor Dip-C has been mapped in D. melanogaster because both loci are apparently monomorphic. Their map positions in D. simulans with respect to flanking markers whose homologous genes have been cytogenetically localized in D. melanogaster suggested that they might be mapped cytogenetically by using available deficiencies in D. melanogaster. Accordingly, by the construction of interspecific hybrids which carried deficiencies of melanogaster and an allele of simulans with a mobility different from that of the fixed melanogaster allele, Dip-B and Dip-C were localized between 87 F12-14 and 88 C1-3 and between 87 B5-6 and 87 B8-10, respectively, in the salivary gland chromosomes of D. melanogaster. The similarity between these two species is discussed on the basis of these findings.

Alleles↗

Molecular evolution of the paired gene in Drosophila: cloning and characterization of the partial paired gene from Drosophila willistoni.

A partial paired gene of Drosophila willistoni containing the paired box and extended homeo box was amplified by PCR and the nucleotide sequence of 1141 bp was determined. Comparison of the paired genes in D. willistoni and D. melanogaster showed that the proportions of identical nucleotide sites in the coding region and identical amino acid sites were 73.8 and 86.5%, respectively. The amino acid sites in the N-terminal region, the paired box, and the extended homeo box were 88.5, 95.3, and 98.6% identical in the two species. The rates of amino acid substitution for these regions were estimated to be 1.73 x 10(-9), 0.67 x 10(-9), and 0.19 x 10(-9)/site/year, respectively. In contrast, the connecting region between the two boxes has been highly diverged and evolved very rapidly, 18.3 x 10(-9)/site/year, suggesting almost no functional constraint in the connecting region.

Amino Acid Sequence↗

Courtship song and mating speed in hybrids between Drosophila melanogaster and Drosophila simulans.

Courtship song and mating speed of hybrids between Drosophila melanogaster and D. simulans were investigated. The courtship song of hybrid males is identical to that of D. simulans, suggesting that X chromosome determination, known from the cross between D. pseudoobscura and D. persimilis, is also possible here. Wingbeat frequency of hybrids is intermediate between that of the two parents, demonstrating that courtship song and wingbeat frequency are inherited independently of each other. In mating test, hybrid males cout and are accepted by D. simulans females more than hybrid females (presumably because their song is more "acceptable" to the former). D. melanogaster males readily, hybrids less readily, and D. simulans least.

Animals↗

Analysis of courtship sequences of the hybrids between Drosophila melanogaster and Drosophila simulans.

Several transitions between sequential male courtship elements were analyzed for Drosophila melanogaster, its close relative D. simulans, and two types of hybrid males. Hybrid males from special reciprocal crosses did not differ. While D. melanogaster and D. simulans males differed markedly for the majority of transitions studied, hybrid males showed no consistent pattern with the parent species, being indistinguishable from D. simulans males, indistinguishable from D. melanogaster, or intermediate between them, depending on the trait observed. This suggests independent genetic control of these transitions during male courtship.

Animals↗

Behavioral role of the sexcombs in Drosophila melanogaster and Drosophila simulans.

The sexcombs were amputated from males of three strains of Drosophila melanogaster and one strain of D. simulans in order to assess the importance of these structures in the sexual behavior of these species. In D. melanogaster the sexcombs are important in attempts to copulate with the female. Their removal delays copulation but does not suppress it entirely. Other aspects of courtship are not influenced by removal of the sexcombs. Strain differences in quanitative aspects of courtship were found, and also in the insemination rates of females by males without sexcombs. The present evidence suggests that the sexcombs are primarily structures adapted to grasping the female securely during the act of intromission.

Animals↗

The Doc transposable element in Drosophila melanogaster and Drosophila simulans: genomic distribution and transcription.

The mobile element Doc is similar in structure and coding potential to the LINE families found in various organisms. In this paper, we analyze the insertional and structural polymorphism of this element and show that it appears to have a long evolutionary history in the genome of D. melanogaster. Like the family of I elements, the Doc family seems to display three types of elements: full length elements, defective members that have recently transposed and long since immobilized members common to each D. melanogaster strain. These three classes of Doc elements seem to be present in D. simulans, a closely related species to D. melanogaster. Furthermore, we show that Doc is transcribed as a polyadenylated RNA of about 5 kb in length, presumed to be a full length RNA. This transcript is present in different tissues and at different stages of Drosophila development. These results are compared with previous records on the chromosomal distribution of LINEs or other transposable element families. Doc transcription is analyzed in an attempt to understand the link between Doc transcription and transposition.

Animals↗

Indirect evidence of alteration in the expression of the rDNA genes in interspecific hybrids between Drosophila melanogaster and Drosophila simulans.

Crosses between Drosophila melanogaster females and D. simulans males produce viable hybrid females, while males are lethal. These males are rescued if they carry the D. simulans Lhr gene. This paper reports that females of the wild-type D. melanogaster population Staket do not produce viable hybrid males when crossed with D. simulans Lhr males, a phenomenon which we designate as the Staket phenotype. The agent responsible for this phenomenon was found to be the Staket X chromosome (Xmel, Stk). Analysis of the Staket phenotype showed that it is suppressed by extra copies of D. melanogaster rDNA genes and that the Xmel, Stk chromosome manifests a weak bobbed phenotype in D. melanogaster Xmel, Stk/0 males. The numbers of functional rDNA genes in Xmel, Stk and Xmel, y w (control) chromosomes were found not to differ significantly. Thus a reduction in rDNA gene number cannot account for the weak bobbed Xmel, Stk phenotype let alone the Staket phenotype. The rRNA precursor molecules transcribed from the Xmel, Stk rDNA genes seem to be correctly processed in both intraspecific (melanogaster) and interspecific (melanogaster-simulans) conditions. It is therefore suggested that the Xmel, Stk rDNA genes are inefficiently transcribed in the melanogaster-simulans hybrids.

Animals↗

Molecular analysis of the intergenic region of the duplicated Amy genes of Drosophila melanogaster and Drosophila teissieri.

The intergenic regions between the duplicated amylase coding regions (Amy) of D. melanogaster and D. teissieri were sequenced. Their lengths in D. melanogaster and D. teissieri were 4,536 bp and 4,621 bp, respectively. Since homology between the upstream regions of the two duplicated genes was found up to 450 bp from the initiation codon of the Amy genes, the ancestral Amy coding region duplicated together with at least 450 bp of the 5'-flanking region as one unit. Comparison of the regions between the two species revealed that the level of divergence was very heterogeneous. Although the mean level of the nucleotide difference in this region was 0.107, no nucleotide substitution was found in four subregions whose sizes were more than 100 bp. Since the probability of these four subregions being completely conserved between D. melanogaster and D. teissieri was very low, these subregions were considered to have relatively important roles in evolution. Large insertions and deletions were not observed in this region but small ones were observed all over the region except for an about 1-kb subregion. This 1-kb region corresponded to an open reading frame encoding a protein which had some sequence identity with the proteins of the serine protease inhibitor superfamily (serpin). Since we could find a transcript of this gene and the synonymous substitution rate was higher than the replacement substitution rate, we suggest that this gene encodes an active serpin in Drosophila.

Amino Acid Sequence↗

Molecular population genetics of the alpha-esterase5 gene locus in original and colonized populations of Drosophila buzzatii and its sibling Drosophila koepferae.

Several studies have suggested that esterase-2 (EST-2) may be the target of natural selection in the cactophilic fly Drosophila buzzatii. In this work, we analyzed nucleotide variation in a fragment of alpha-esterase5 (alphaE5), the gene encoding EST-2, in original (Argentinian) and colonized (Australian) populations of D. buzzatii and in its sibling D. koepferae. Estimates of nucleotide heterozygosity in D. buzzatii were similar in Australia and Argentina, although we detected a loss of singletons in colonized populations, suggesting a moderate founder effect. Interspecific comparisons revealed that D. buzzatii was more polymorphic for nonsynonymous variation, whereas D. koepferae was more variable for synonymous and noncoding sites. The two major chromosomal arrangements (2st and 2j) in D. buzzatii displayed similar levels of nucleotide variation, whereas 2jz3 was monomorphic. The sequenced region allowed the discrimination of a greater number of EST-2 protein variants in the Australian sample than in the Argentinean sample. In D. koepferae, nucleotide variation in alphaE5 does not depart from neutral expectations, although tests of population structure were significant for silent variation. In contrast, D. buzzatii has probably undergone a recent population expansion in its South American range. In addition, the McDonald and Kreitman test revealed an excess of nonsynonymous polymorphism in both original and colonized populations of this species.

Animals↗

Comparison of the GAGA factor genes of Drosophila melanogaster and Drosophila virilis reveals high conservation of GAGA factor structure beyond the BTB/POZ and DNA-binding domains.

As a member of the trithorax-group, the Trithorax-like (Trl) gene of Drosophila melanogaster contributes to the expression of homeotic genes and many other genes. Trl encodes different isoforms of the GAGA factor which is thought to act as an "antirepressor" of transcription by remodelling chromatin structure and thereby rendering control regions accessible for transcriptional activators. A more global role of the GAGA factor in chromatin structure and function is suggested by various phenotypes of Trl mutations, such as modification of position effect variegation. To better define the molecular basis of these pleiotropic effects, we cloned cDNAs encoding the GAGA isoforms of D. melanogaster and a distantly related species, D. virilis. We also characterized the genomic organization of both the D. melanogaster and D. virilis genes, and analysed the expression patterns of isoform-specific mRNAs. The D. virilis GAGA isoforms show high similarity to their D. melanogaster counterparts, particularly within the BTB/POZ protein-interaction and the zinc finger DNA-binding domains. Interestingly, conservation clearly extends beyond the previously defined limits of these domains. Moreover, the comparison reveals a completely conserved block of amino acid residues located between the BTB/POZ and DNA-binding domains, and a high conservation of the C-terminus specific for one of the GAGA isoforms. Thus, sequences of as yet unknown functions are defined as rewarding targets for further mutational analyses. The high conservation of the GAGA proteins of the two species is in accord with the nearly identical genomic organization and expression patterns of the corresponding genes.

Amino Acid Sequence↗

Molecular genetics of the Alhambra (Drosophila AF10) complex locus of Drosophila.

The Alhambra (Alh) gene is the Drosophila homologue of the human AF10 gene. AF10 has been identified as a fusion partner of MLL, a human homologue of the fly gene trithorax, in infant leukemias. The endogenous function of human AF10 is not known, but may be vital to its role in acute leukemia. This prompted us to analyse Alh function. We describe here the genetic organisation of the Alh locus in D. melanogaster. We show that an independent lethal complementation group encoding a muscle protein (Mlp84B) is located within an Alh intron. We have already shown that the leucine zipper (LZ) domain of ALH activates several Polycomb group-responsive elements. We further demonstrate that the LZ domain on its own bears the Alh vital function, since it is necessary and sufficient for rescue of Alh mutant lethality. Finally, we demonstrate that, in contrast to a previous report, Alh does not affect position-effect variegation.

Animals↗

The endogenous Drosophila melanogaster retrovirus gypsy can propagate in Drosophila hydei cells.

The endogenous Drosophila melanogaster retrovirus gypsy (mdg4) forms virus-like particles (VLPs) which are found as extracellular particles in the medium used to culture D. melanogaster cells. The D. hydei somatic cell line DH14, which does not harbour gypsy sequences, was exposed to D. melanogaster VLPs. Subsequent PCR and Southern analysis revealed that gypsy elements had penetrated into the D. hydei cells, suggesting interspecific transmission of the retrovirus. A D. hydei cell line containing gypsy sequences was established and grown in a mixed culture together with the G418-resistant D. hydei cell line DH33, and gypsy was shown to be transmitted from cell to cell. The proportion of cells carrying gypsy increased with time. The rate of gypsy invasion of the lines DH14 and DH33 was 10(-3) and 10(-2) per cell per generation, respectively. The results demonstrate the possibility of interspecific horizontal transfer of gypsy in the form of its VLPs.

Animals↗

Complex developmental regulation of the Drosophila affinidisjuncta alcohol dehydrogenase gene in Drosophila melanogaster.

During development, the alcohol dehydrogenase genes of Drosophila melanogaster and D. affinidisjuncta are expressed in similar, yet distinct, tissue- and stage-specific patterns. Transcripts from both of these genes arise from two promoters (distal and proximal) that also display tissue and stage specificity. We used P-element-mediated transformation to introduce the D. affinidisjuncta Adh gene into the germ line of D. melanogaster. We show that the D. affinidisjuncta Adh gene is expressed at comparable overall levels in both species and that the tissue- and stage-specific expression for this gene (including promoter utilization) is similar in the donor and the host species. However, in some details, the expression of the D. affinidisjuncta gene in D. melanogaster resembles the host pattern, and one novel tissue-specific expression phenotype is displayed by transformants. In general, our results suggest that there has been strong conservation of cis- and trans-acting regulatory factors since the divergence of the two species but that this conservation has not been perfect.

Alcohol Dehydrogenase↗

Positive and negative DNA elements of the Drosophila grimshawi s18 chorion gene assayed in Drosophila melanogaster.

Germ line transformation has been used to map the cis regulatory DNA elements responsible for the precise and evolutionarily stable developmental expression of the s18 chorion gene. Constructs containing chimeric combinations of Drosophila melanogaster and D. grimshawi DNA regions, as well as D. grimshawi sequences alone, can direct expression in the follicular epithelium, in an s18-specific temporal and spatial pattern. The results indicate that both positive and negative regulatory elements can function when transferred from D. grimshawi to D. melanogaster. The first ca. 100 bp of the 5'-flanking DNA region constitute a minimal, developmentally regulated promoter, expression of which is inhibited by the next 100-bp DNA segment and activated by positive elements located further upstream. Expression of the minimal promoter can also be enhanced by more distant chorion regulatory elements, provided the inhibitory DNA segment is absent.

Animals↗

Structure of Drosophila virilis glycerol-3-phosphate dehydrogenase gene and a comparison with the Drosophila melanogaster gene.

The complete glycerol-3-phosphate dehydrogenase gene of Drosophila virilis isolated by screening with alpha GPDHM cDNA of the adult fly was sequenced. The gene contains eight exons spread over a total of approximate 8 kb DNA. Its exon/intron organization is identical to that of D. melanogaster. A single transcription initiation site was determined by primer extension. The stop codons are located at the 3' end of each of the exons 6 to 8. TATA and CAAT boxes are present upstream of the transcriptional start site. Adult alpha GPDH protein is encoded by exons 1 to 6 and exon 8. Comparison of the sequence with that of D. melanogaster showed that the homology of the nucleotide sequence of the coding region is 85% and that the homology of the amino acid sequence is 98%. On the contrary, the non-coding region is quite different in length and nucleotide sequence.

Animals↗

The Drosophila melanogaster similar bHLH-PAS gene encodes a protein related to human hypoxia-inducible factor 1 alpha and Drosophila single-minded.

The Drosophila melanogaster (Dm) similar (sima) gene was isolated using a low-stringency hybridization screen employing a Dm single-minded gene basic helix-loop-helix (bHLH) DNA probe. sima is a member of the bHLH-PAS gene family and the conceptual protein shares a number of structural features, including a bHLH domain, PAS domain, and homopolymeric amino acid stretches. Sima is most closely related to the human hypoxia-inducible factor 1 alpha bHLH-PAS protein. In situ hybridization experiments reveal that sima is transcribed in most or all cells throughout embryogenesis. It has been cytologically mapped to position 99D on the third chromosome, and is not closely linked to other known bHLH-PAS genes.

Amino Acid Sequence↗

Conservation of structure and expression of the trithorax gene between Drosophila virilis and Drosophila melanogaster.

The Drosophila melanogaster trithorax gene encodes several large RNAs which are expressed in complex patterns in the embryo. The D. virilis trithorax gene was isolated and sequenced. It produces a similar to D. melanogaster set of transcripts, and it encodes a protein that shows sequence similarity in several domains which are also conserved in human homologue, ALL-1/HRX. Previous experiments have suggested that a distinct expression domain of trithorax in the posterior region of the embryo is required to maintain expression of the BX-C genes (Sedkov et al., 1994, Development 120, 1907-1917). At cellular blastoderm, trithorax RNA expression in D. virilis embryos is also confined to the posterior portion of the presumptive mesoderm. This finding supports the idea that the specific BX-C-related expression domain is an essential feature of the trithorax gene.

Amino Acid Sequence↗