Search PubMed⌕ Search

Biomedical subjects

W Hennig

Publications and source records attributed to W Hennig.

At least 37 records · Page 2Linked to original sources
PubMed1994

Degenerating gypsy retrotransposons in a male fertility gene on the Y chromosome of Drosophila hydei.

During the evolution of the Y chromosome of Drosophila hydei, retrotransposons became incorporated into the lampbrush loop pairs formed by several of the male fertility genes on this chromosome. Although insertions of retrotransposons are involved in many spontaneous mutations, they do not affect the functions of these genes. We have sequenced gypsy elements that are expressed as constituents of male fertility gene Q in the lampbrush loop pair Nooses. We find that these gypsy elements are all truncated and specifically lost those sequences that may interfere with the continuity of lampbrush loop transcription. Only defective coding regions are found within the loop. Gypsy is not transcribed in loops of many other Drosophila species harboring the family. These results suggest that any contribution of gypsy to the function of male fertility gene Q does not depend on a conserved DNA sequence.

Animals↗
PubMed1994

Discrimination of related transcribed and non-transcribed repetitive DNA sequences from the Y chromosomes of Drosophila hydei and Drosophila eohydei.

The short arm of the Y chromosome of Drosophila hydei carries a single male fertility gene, gene Q, which forms the lampbrush loop pair Nooses. Conflicting observations have been reported concerning the identity of the repetitive DNA sequences that are transcribed in this loop pair. It has been claimed by other investigators that the loop transcripts contain repeats of two distinct, but related families of Y-specific repetitive DNA sequences, ay1 and YsI. We reinvestigated this issue, using as probes single ay1 and YsI repeats which, under stringent conditions, hybridize only to members of their own family. Under non-stringent conditions, both repeats hybridize in situ to Nooses transcripts. However, if hybridization conditions are stringent, only the ay1 probe hybridizes to loop transcripts. Hybridizations to Northern blots of testis RNA confirm these results. Further, YsI repeats are not found the closely related species D. eohydei. We conclude that the YsI repeats are not relevant for the function of fertility gene Q.

Animals↗
PubMed1993

Partial reconstruction of the lampbrush loop pair Nooses on the Y chromosome of Drosophila hydei.

We present the analysis of genomic DNA fragments that were isolated as potential segments of the lampbrush loop pair Nooses on the short arm of the Y chromosome of Drosophila hydei. More than 300 kb of DNA were recovered in BamHI lambda and cosmid clone groups. This DNA is composed of the Y-specific ay1 family of repetitive DNA sequences, and of other repetitive DNA sequences, which at least in part are also located elsewhere in the genome (Y-associated sequences). Two additional classes of DNA fragments were obtained from an EcoRI library. One of them consists of ay1 repeats without apparent interspersion, including a total of more than 300 kb of DNA. The other is composed of tandemly repeated YsI sequences, a Y-specific sequence derived from ay1. This class includes more than 400 kb of DNA, which is also not interspersed by other sequences. Our results show that only the ay1 repeats interspersed by Y-associated DNA sequences can represent parts of the 260 kb transcription unit forming the lampbrush loop, whereas the ay1 and YsI repeats without interspersion form separate and nontranscribed clusters of repetitive DNA.

Animals↗
PubMed1993

Localization of the lampbrush loop pair Nooses on the Y chromosome of Drosophila hydei by fluorescence in situ hybridization.

We have used fluorescence in situ hybridization to map the positions of the different repetitive DNA sequences from the region forming the lampbrush loop pair Nooses on the Y chromosome of Drosophila hydei. This region harbours a megabase cluster of tandemly organized repeats of the Y-specific ay1 family and a megabase cluster of tandem repeats of the related Y-specific YsI family. In addition, ay1 repeats also occur in short blocks that are interspersed by other repetitive DNA sequences that we call Y-associated, since they have additional copies on other chromosomes. Using specific probes for ay1, YsI and Y-associated DNA sequences, we show that there is one large proximal cluster of YsI repeats and one, more distally located, large cluster of ay1 repeats. The Y-chromosomal copies of the Y-associated sequences are located in the most distal part of the ay1 cluster. This is consistent with the juxtaposition of ay1 and Y-associated sequences in more than 300 kb of cloned genomic DNA. Since both ay1 and Y-associated sequences have been shown to be transcribed in the Nooses, the lampbrush loop is formed in a distal region of the short arm of the Y chromosome, adjacent to the terminally located nucleolus organizer region. The clusters of homogeneous ay1 and YsI repeats are of no functional significance for the formation of the lampbrush loop.

Animals↗
PubMed1992

Laminin in the male germ cells of Drosophila.

To study genes that may be crucial for the male germ cell development of Drosophila we screened a cDNA expression library with a polyclonal antiserum against testis proteins of Drosophila hydei. We identified a cDNA fragment that exhibited a complete sequence similarity with the cDNA of the laminin B2 chain, an important component of the extracellular matrix. Transcripts of laminin B2 were detected in the RNA of male germ cells with the polymerase chain reaction and by in situ hybridization. We studied the reaction of different polyclonal antibodies including those against a Drosophila laminin B2-lac fusion protein, the entire Drosophila laminin complex, or against the mouse laminin complex and against laminin A and B1 chains with specific structures in developing male germ cells of Drosophila. Antigenic sites against laminin B2 were found in the lampbrush loops in primary spermatocyte nuclei, in nuclei of spermatids, and in heads of spermatozoa. The axonemes of elongating spermatids react with antibodies against the Drosophila laminin B1, B2 and laminin A chains. The possible biological functions of the laminin in the male germ cells of Drosophila are discussed.

Animals↗
PubMed1990

Isolation and characterization of a Drosophila hydei histone DNA repeat unit.

Histone genes in D. hydei are organized in tandemly repeated clusters., accomodating in total 120-140 repeat units. We cloned one of the repeat units and analysed the nucleotide sequence. The repeat unit has a size of 5.1 x 10(3) base-pairs and contains one copy of each of the genes coding for the core histones and one copy coding for the histone H1. In the promoter regions of the genes we identified the presumptive cap sites and TATA boxes. Two additional sequence elements are shared by all five Drosophila hydei histone genes in the cluster. The sequence CCCTCT/G1 is found in the region upstream of the presumptive CAP sites. The sequence element AGTGAA occurs downstream of the presumptive cap sites and is, in contrast to the promoter element, also seen in the histone genes of Drosophila melanogaster. Cell-cycle dependent regulation of transcription of the Drosophila histone genes may be different from that in other eukaryotes since sequence elements involved in the regulation of cell-cycle dependent transcription are absent. Also other regulatory elements for transcription differ from those of other genes. The highly conserved H1-specific promoter sequence AAACACA and the H2B specific promoter sequence ATTTGCAT, which are involved in the cell-cycle dependent transcription of those histone genes in eukaryotes, are missing in the Drosophila genes. However at the 3' end of the genes the palindrome and the purine-rich region, both conserved sequence elements in histone genes of eukaryotes, are present. The spacer regions show a simple sequence organization. The silent site substitution rate between the coding regions of the D. hydei and D. melanogaster histone genes is at least 1.5 times higher for Drosophila than for sea urchin histone genes.

Amino Acid Sequence↗
PubMed1990

Poly[d(C-A)].poly[d(G-T)] is highly transcribed in the testes of Drosophila hydei.

Microdissection of the lampbrush loops "threads" and "pseudonucleolus" of Y chromosomes from primary spermatocytes of Drosophila hydei and subsequent microcloning of the DNA yielded several recombinant DNA clones which cross-hybridized in screening the different clone banks. By DNA sequencing we found that the inserts of these cross-hybridizing clones contain blocks of poly[d(C-A].poly[d(G-T)]. Testis RNA contains a large fraction of transcripts with this simple repeated nucleotide sequence. With the aid of transcript in situ hybridization we discovered that the "cones" and "pseudonucleolus" lampbrush loops are the primary sites of transcription of poly[d(C-A)].poly[d(G-T)] in spermatocytes. In addition, we found a strand-specific transcription of (CA/GT)n. In both the "cones" and "pseudonucleolus" the (CA)n strand is transcribed, while in the "pseudonucleolus" (GT)n is also transcribed. Labelled (CA)n probes also react with the protein bodies in spermatid nuclei. These observations are discussed in the context of possible functions of (CA/GT)n transcripts in spermatogenesis.

Animals↗
PubMed1990

DNA sequence comparison of micropia transposable elements from Drosophila hydei and Drosophila melanogaster.

Members of the retrotransposon family micropia were discovered as constituents of wild-type Y chromosomal fertility genes from Drosophila hydei. Several members of the micropia family have subsequently been recovered from Drosophila melanogaster and four micropia elements, micropia-DhMiF2, -DhMiF8, -Dm11 and -Dm2, two each from D. hydei and D. melanogaster, have been totally sequenced (17 kb of micropia sequences and 6.8 kb from insertions). Comparative analysis of micropia sequences revealed a complex pattern of divergence within a single Drosophila genome. The divergence includes deletions, possibly by a slipped mispairing mechanism, insertions of a retroposon, and of another retrotransposon (copia) and "positional nucleotide shuffling" within the tandem repeats of the 3' non-protein-coding region of micropia elements. A 10 bp long sequence of each repeat unit of the 3' tandem repeats of micropia elements is highly conserved and is therefore a candidate of functional importance either in transposition events or in regulatory activity on flanking DNA sequences.

Animals↗
PubMed1989

Characterization of the long terminal repeats of micropia elements microdissected from the Y-chromosomal lampbrush loops "threads" of Drosophila hydei.

Four micropia elements from Drosophila melanogaster and D. hydei have been analysed by sequencing. Two elements, from D. hydei, micropia-DhMiF8 and -DhMiF2, were recovered by cloning microdissected Y-chromosomal lampbrush loops "threads". This method allows isolation of repetitive sequences from defined chromosomal positions, but recovery of large and overlapping inserts is difficult. In case of the Y-chromosomal micropia elements it was not possible to define the endpoints of their long terminal repeat sequences precisely. Comparison of these locus-defined micropia elements to complete micropia elements isolated from D. melanogaster allowed identification of micropia-DhMiF8 and micropia-DhMiF2 long terminal repeats (LTRs). LTR sequences from the two Drosophila species are not conserved except for a few short sequences found at comparable positions that are believed to have functional significance. In contrast, the Leu-tRNA primer binding site and plus strand primer binding site are conserved between D. melanogaster and D. hydei.

Animals↗
PubMed1989

An abundant testis RNA species shows sequence similarity to Y chromosomal and other genomic sites in Drosophila hydei.

A cDNA clone bank was constructed from testis poly(A)+ RNA of Drosophila hydei and screened for clones which hybridize to Y chromosomal DNA sequences. The insert of clone cDhT14 hybridizes to a family of repeated DNA sequences with members distributed within the Y chromosome and elsewhere in the genome. This type of sequence has earlier been described as the Y-associated class of DNA. Southern blot analysis of DNA from different wild-type strains of D. hydei suggests that members of the T14 family of repeated DNA sequences are parts of a family of transposable elements. The genomic localization of the T14 family of repeated DNA sequences was revealed by in situ hybridization to metaphase and polytene chromosomes, and to transcripts of Y chromosomal lampbrush loops. Approximately 10-15 members (20%-30%) of the T14 sequence family reside in 8.3 kb PstI restriction fragments. A genomic clone of one of these DNA fragments, DhT14-8.3, hybridizes to transcripts on the Y chromosomal lampbrush loop "cones", and in conventional in situ hybridization experiments to region 12D/13A of the X chromosome and to region 112 of chromosome 5. The cDNA clone cDhT14 represents a part of an abundant testis RNA species of 5.0 kb. This RNA is also present in ovaries and in 0-3 h, 3-6 h and 6-12 h embryos, but less abundantly than in testes. Both the Y chromosomal site of the 8.3 kb PstI fragments and sites elsewhere in the genome are actively transcribed. At least one of the latter genomic sites is transcribed into the 5.0 kb RNA species.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1989

Y chromosomal fertility genes of Drosophila: a new type of eukaryotic genes.

The Y chromosomal fertility genes of Drosophila are required for sperm differentiation. They are active only in primary spermatocytes where they form giant lampbrush loops. The molecular structure of these genes was investigated and revealed an unusual composition of DNA. Short, tandemly repeated sequence clusters are interrupted by longer and more heterogeneous sequences, which probably all represent transposable elements. No indication of the presence of protein-coding regions has been found within the fertility genes. However, the lampbrush loops bind site-specific proteins recognized by immunofluorescence techniques. This, together with other experimental data, led to the hypothesis that the Y chromosomal genes have a function in binding chromosomal proteins. The data and arguments in support of this gene model are summarized in this paper.

Animals↗
PubMed1988

Micropia: a retrotransposon of Drosophila combining structural features of DNA viruses, retroviruses and non-viral transposable elements.

The retrotransposon micropia was first described from Y-chromosomal fertility genes of Drosophila hydei. Screening a Drosophila melanogaster genomic library yielded several clones representing micropia elements in D. melanogaster. The DNA sequences of two elements from D. hydei (micropia-DhMiF2 and micropia-DhMiF8) and two elements from D. melanogaster (micropia-Dm2 and micropia-Dm11) permitted a detailed analysis of the spatial organization of micropia constituents. Micropia represents the typical gene organization represented by "core"-protein domains followed by a protease, reverse transcriptase, RNase and integrase domain. New features of the micropia family compared with other retrotransposons are: (1) a region of similarity to class I major histocompatibility complex antigens of mammals; (2) only one main open reading frame of about 4000 bases length; (3) a non-protein-coding region of about 500 base-pairs length between the 3' end of the open reading frame and the 5' start of the 3' long terminal repeat. This region includes 32 base-pair tandem repeats; (4) within the long terminal repeats, 82 base-pair tandem repeats with four potential ecdysteroid receptor binding sites. Because micropia combines many evolutionary features of different viruses, non-viral transposable elements, chromosomal genes and repetitive sequence organizations, this retrotransposon may be seen as a "minigenome" reflecting evolutionary principles of the construction of genomic components.

Amino Acid Sequence↗
PubMed1988

Retrotransposon-like sequences are expressed in Y chromosomal lampbrush loops of Drosophila hydei.

The DNA sequence family micropia consists of repeated DNA sequences that occur dispersed in the genome of Drosophila hydei. Members of this DNA sequence family were recovered from two recombinant DNA clone banks obtained by microdissection of the two Y chromosomal lampbrush loop threads and pseudonucleolus from primary spermatocyte nuclei. Nucleotide sequence analysis of two of the recombinant DNA clones revealed homology to the DNA region coding for a reverse transcriptase-like protein in retroviruses and retrotransposons. Homologous tissue-specific transcripts of a size of 1.2 x 10(3) base-pairs were found in testes. Transcript in-situ hybridization shows that at least parts of these transcripts are synthesized in these Y chromosomal lampbrush loops, which were originally used for microdissection. Also the cytoplasm of primary spermatocytes contains homologous RNA species. These observations are discussed in the context of lampbrush loop function and evolution.

Amino Acid Sequence↗