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M Frasch

Publications and source records attributed to M Frasch.

52 records · Page 3Linked to original sources
PubMed1991

Mutation of the hamster cell cycle gene RCC1 is complemented by the homologous genes of Drosophila and S.cerevisiae.

The RCC1 gene has been isolated from several vertebrates, including human, hamster and Xenopus. Genes similar to RCC1, namely BJ1 and SRM1/PRP20, have been isolated from the insect Drosophila and from the budding yeast Saccharomyces cerevisiae. A mutation of the RCC1 gene in the hamster BHK21 cell line, tsBN2, confers pleiotropic phenotypes, including G1 arrest and premature induction of mitosis in cells synchronized at the G1/S boundary. Similarly, mutations of the SRM1/PRP20 gene are pleiotropic; the srm1 mutant shows G1 arrest and suppression of the mating defect of mutants lacking pheromone receptors, and the prp20 mutant shows an alteration in mRNA metabolism. Here we show that both BJ1 and SRM1/PRP20 complement the temperature sensitive phenotype of the tsBN2 cells. Like RCC1 proteins of vertebrates, the protein products of the Drosophila and yeast RCC1 homologues were located in the nuclei of the mammalian cells. These results suggest that the BJ1 and SRM1/PRP20 genes are functionally equivalent to the vertebrate RCC1 genes, and that the RCC1 gene plays an important role in the regulation of gene expression in the eukaryotic cell cycle.

Amino Acid Sequence↗
PubMed1991

Characterization of a Drosophila protein associated with boundaries of transcriptionally active chromatin.

We have used indirect immunofluorescence of polytene chromosomes to examine the chromatin distribution of a 52-kD Drosophila protein designated B52. B52 is localized to transcriptionally active loci and, at the highly decondensed heat shock loci, can be seen to bracket the RNA polymerase II fluorescence signals symmetrically. We have also examined the distribution of B52 on nonpolytene chromosomes in Drosophila cell cultures with an in vivo UV cross-linking method and find that, here too, B52 is associated with boundaries of transcriptionally active chromatin. The predicted primary amino acid sequence of B52 reveals two regions with similarities to a number of other proteins known to interact with nucleic acids.

Amino Acid Sequence↗
PubMed1990

Two puff-specific proteins bind within the 2.5 kb upstream region of the Drosophila melanogaster Sgs-4 gene.

The Drosophila nuclear proteins Bj6 and Bx42 characterized previously are detected in a series of developmentally active puffs on salivary gland chromosomes. Here the binding of both proteins at puff 3C11-12 containing the glue protein gene Sgs-4 is described in more detail. By deletion analysis we show that both proteins bind within a chromosomal segment containing 17-19 kb of DNA surrounding the Sgs-4 gene. They are detectable at this site during the intermoult stages, before the puff regresses in response to the moulting hormone ecdysone. If the Sgs-4 gene together with flanking DNA sequences is brought into a different chromosomal position by P element transfer, both proteins are detected at this new location. Both proteins are bound to the chromosome within the range of 2.5 kb DNA upstream of the Sgs-4 gene. A strain containing a 52 bp deletion within this region fails to bind Bx42 protein suggesting that the missing DNA, which overlaps a hypersensitive region, may be required for the binding of the Bx42 protein.

Animals↗
PubMed1990

A new Drosophila homeo box gene is expressed in mesodermal precursor cells of distinct muscles during embryogenesis.

Several Drosophila homeo box genes have been shown to control cell fates in specific positions or cell groups of the embryo. Because the mechanisms involved in the pattern formation of complex internal organs, such as the musculature and the nervous system, are still largely unknown, we sought to identify and analyze new homeo box genes specifically expressed in these tissues. Here, the molecular analysis and expression pattern of one such gene, containing both a homeo box and a PRD repeat, is described. This gene, designated S59, is expressed in a small number of segmentally repeated mesodermal cells approximately 2 hr postgastrulation. Gradually, four groups of S59-expressing mesodermal cells appear in each abdominal hemisegment, each one giving rise to a particular somatic muscle after fusion with surrounding myoblasts. Thus, individual precursors for particular muscles, which we call "founder cells," are specified relatively early during mesodermal development. The expression of a particular homeo box gene in these cells suggests that distinct programs of gene expression are active in subsets of mesodermal cells after germ band elongation, resulting in a specification of their developmental fates. In addition to the mesoderm, S59 is expressed in a subset of neuronal cells of the CNS and their precursors and also in cells of a small region of the midgut.

Amino Acid Sequence↗
PubMed1989

Two proteins from Drosophila nuclei are bound to chromatin and are detected in a series of puffs on polytene chromosomes.

Immunizing chromatin protein fractions from Drosophila melanogaster embryos, monoclonal antibodies have been generated against two nuclear proteins of different molecular weight. These proteins are present in a chromatin fraction of Drosophila Kc-cell nuclei and both proteins could be shown to cosediment with nucleosomes following separation on sucrose gradients. Early in development both proteins are located in the embryo cytoplasm. Later, at times when transcription starts at blastoderm, they become redistributed into the nuclei. On salivary gland chromosomes both proteins are detected in a series of developmentally active puffs. The number of sites where these antigens can be detected, as well as the qualitative properties of the antigens at these sites differ between both proteins.

Animals↗
PubMed1988

Molecular analysis of even-skipped mutants in Drosophila development.

The homeo box gene even-skipped (eve) plays a key role in the regulation of the Drosophila segmentation pattern. eve- embryos lack segment borders and show altered activities of several segmentation genes, including fushi tarazu (ftz), engrailed (en), and wingless (wg). Here, we present evidence that eve influences its own expression in a tissue-specific manner. Each of four different eve mutations disrupts the normal eve expression pattern, and null mutations cause a premature loss of eve products in ectodermal, but not mesodermal, tissues. Molecular characterization of eve mutations indicates that disruptions of the eve pattern are not due to alterations in the eve promoter but, instead, involve abnormal eve proteins. Two different eve mutations cause single amino acid substitutions within the homeo box, and we discuss the implications of these changes with regard to homeo box gene function. We also present evidence that eve+ gene activity is not only required for the activation of the odd-numbered en stripes but also for the correct positioning of each ftz stripe. We present a model for the loss of en expression in eve- embryos, based on the concentration-dependent regulation of the ftz pattern by eve+ products.

Alleles↗
PubMed1988

Cloning of a gene encoding an antigen associated with the centrosome in Drosophila.

The monoclonal antibody Bx63 recognizes a centrosomal antigen of Drosophila melanogaster by indirect immunofluorescence and identifies two proteins, with apparent molecular weights of 185 x 10(3) and 66 x 10(3), on Western blots. We have used this antibody to isolate five clones (lambda cs1, -2, -3, -4 and lambda j63) from lambda gt11 expression libraries of Drosophila DNA. Using polyclonal anti-centrosomal sera raised against both immunoaffinity-purified Bx63 antigen and electrophoretically purified fusion protein from clone lambda cs3, we have demonstrated that the fusion proteins encoded by four of these clones (lambda cs1-4) share at least two epitopes with the 185 x 10(3) Mr centrosomal antigen. This indicates that clones lambda cs1-4 contain DNA from the gene coding for this protein. The four clones are independent isolates from a single chromosomal site, which we show by in situ hybridization to correspond with salivary gland chromosome region 88E 4-8. A low-abundance transcript of approximately 4.0 x 10(3) bases corresponding to the cloned gene is detected in all stages of the Drosophila life-cycle.

Animals↗
PubMed1988

Developmental and mitotic behaviour of two novel groups of nuclear envelope antigens of Drosophila melanogaster.

Two novel groups of nuclear envelope antigens have been identified using monoclonal antibodies. On immunoblots the antigens correspond to distinct sets of polypeptides in the 175 X 10(3) molecular weight range. The antigens are enriched in a nuclear matrix-pore complex-lamina fraction of Drosophila tissue culture cells. We have studied the cellular distribution of these antigens throughout oogenesis and early embryo development. Immunoblots show that one group of the 175 X 10(3) Mr antigens is maternally transmitted to the embryo. This had already been observed for the 74/76 X 10(3) Mr Drosophila lamins described previously, and we showed that a large proportion of the lamins is localized in the interior of the oocyte nucleus. We have also followed the fate of the high molecular weight antigens during mitosis. Each of the antigens uses a different pathway for its distribution to the daughter nuclei. These observations may give clues to the molecular mechanisms involved in the disassembly-reassembly process of the nuclear envelope.

Animals↗
PubMed1987

Characterization and localization of the even-skipped protein of Drosophila.

On the basis of homeo box cross-homology we have isolated the pair-rule gene even-skipped (eve) of Drosophila. The eve transcription unit appears to be less than 1.5 kb in length, and encodes a single mRNA of approximately 1.4 kb. The nucleotide sequence of genomic and cDNA clones indicates that the eve protein is composed of 376 amino acid residues, and that its homeo domain shares only approximately 50% amino acid identity with the homeo domains of previously characterized genes. Using antibodies raised against a beta-galactosidase fusion protein we show that the eve protein is distributed in a series of seven transverse stripes at the cellular blastoderm stage, and is localized primarily within the nuclear regions of those embryonic cells that express the gene. After gastrulation, seven weakly stained stripes of eve expression appear, resulting in a transient pattern that consists of a total of 14 evenly spaced stripes. Both the original and new stripes gradually disappear during germ band elongation. A second expression pattern emerges during neurogenesis, whereby eve protein is detected in discrete subsets of neurons in each of the ventral ganglia.

Amino Acid Sequence↗
PubMed1987

Complementary patterns of even-skipped and fushi tarazu expression involve their differential regulation by a common set of segmentation genes in Drosophila.

We have examined the position-dependent expression of two segmentation genes that contain a homeo box, even-skipped (eve) and fushi tarazu (ftz). Products encoded by both genes accumulate in a series of seven transverse stripes along the anterior-posterior body axis of developing embryos. Double-staining experiments show that eve and ftz proteins accumulate in complementary sets of embryonic cells. A total of eight zygotically active segmentation genes are required for the establishment and refinement of the eve and ftz expression patterns during development. Mutations that affect one of the genes were always found to produce a reciprocal alteration in the other. These results suggest that the eve and ftz promoters independently "interpret" the same positional cues to give differential patterns of expression. We propose that a combination of segmentation gene products that activates the expression of eve represses the expression of ftz, and vice versa. This proposal is discussed in the context of a hierarchy of interactions among segmentation genes, whereby the gap genes establish asymmetric patterns of eve and ftz expression in blastoderm-stage embryos, and interactions with other pair-rule genes serve to refine further the complementarity of eve and ftz expression during gastrulation and germ band elongation.

Animals↗
PubMed1986

Nuclear antigens follow different pathways into daughter nuclei during mitosis in early Drosophila embryos.

In the early embryonic development of Drosophila melanogaster, there is a series of 13 rapid and highly synchronous nuclear divisions. We have used a collection of monoclonal antibodies to follow the re-distribution of nuclear antigens into daughter nuclei at this developmental stage by indirect immunofluorescence microscopy. The antigens fall into several categories in terms of the pathways that are followed at mitosis. At one extreme is a group of antigens that remains continuously associated with the DNA throughout all the mitotic phases. At the other extreme, another group of antigens is excluded from the nucleus at prophase, and does not associate with the nucleus again until late telophase. One antigen, which becomes incorporated into the nucleolus at cellularization after the thirteenth division, becomes associated with the chromosomes during mitosis, but not until anaphase. Several different antibodies stain a diamond-shaped compartment that develops over the spindle at anaphase. The distribution of antigens within this spindle compartment shows some variation: one antigen appears to be present at higher concentrations in the central region of the spindle; others appear in three quite distinct areas corresponding to the positions of the new daughter nuclei and the old parental nucleus. Yet another antibody gives uniform staining of the spindle compartment. This antibody also recognizes a protein present in centrosomes.

Animals↗
PubMed1983

Nonpackaging and packaging proteins of hnRNA in Drosophila melanogaster.

Monoclonal antibodies have previously been raised against chromosomal proteins of Drosophila. Using a biochemical fractionation method for the isolation of large hnRNA-containing structures (hnRNP) of Drosophila tissue culture cells, we show that seven of these antibodies recognize different antigens, and that these antigens are associated with RNA. Analysis of the sedimentation behavior of antigen-containing structures in sucrose gradients reveals that the antigens are differentially distributed with respect both to one another and to pulse-labeled RNA. We demonstrate that the antigens are minor components of hnRNP and are different from the major Drosophila hnRNP packaging proteins, which we have also identified. The antigens are probably involved in the processing of hnRNA in the nucleus.

Animals↗
PubMedDate not supplied

Maternal regulation of zerknüllt: a homoeobox gene controlling differentiation of dorsal tissues in Drosophila.

The homoeobox gene zerknüllt (zen) plays an important role in the differentiation of dorsal tissues during Drosophila development. zen- embryos show transformations in the dorsal-most regions of the fate map, and lack several tissues that normally derive from these regions, including the amnioserosa and optic lobe. zen displays a simple dorsal on/ventral off pattern as early as cleavage cycle 10-11 (ref. 2). We have prepared a polyclonal antibody against a full-length zen protein, and used this to examine its pattern of expression in mutants that disrupt dorsal-ventral polarity. Most or all of the maternally expressed genes that are involved in this process have been previously identified and fall into two classes, so called 'dorsalizers' and 'ventralizers' (see refs 4-7, reviewed in ref. 8). On the basis of our analysis of zen expression in each of these maternal mutants we propose that one or more of the dorsalizing genes encodes a repressor which inhibits the expression of zen in ventral regions of developing embryos. The ventralizing gene cactus might play an important role in restricting the activity of this repressor to ventral regions, thereby permitting the activation of zen in those dorsal tissues where its function is critically required.

Animals↗