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M Bienz

Publications and source records attributed to M Bienz.

At least 73 records · Page 4Linked to original sources
PubMed1989

An essential role of even-skipped for homeotic gene expression in the Drosophila visceral mesoderm.

We have analysed homeotic gene expression in the embryonic visceral mesoderm of segmentation mutants by antibody staining against Ultrabithorax, Antennapedia and Sex combs reduced protein. We found that even-skipped (eve) function is crucially required for homeotic gene expression, whereas most other segmentation mutations have only minor effects on position and/or width of the homeotic expression domains in this germ layer. Analysis of pair-rule double mutants indicates that complete loss of homeotic gene activity in the visceral mesoderm, as observed in amorphic eve mutants, correlates with loss of engrailed (en) expression in the epidermis and loss of segmentation. We suggest that the establishment of parasegment borders, a consequence of eve expression and witnessed by subsequent en expression, is a necessary precondition for homeotic gene expression in the visceral mesoderm.

Animals↗
PubMed1988

Drosophila homoeotic genes encode transcriptional activators similar to mammalian OTF-2.

Homoeotic genes in Drosophila melanogaster are active in spatially restricted metameric domains and control the morphogenesis of segment-specific features such as legs or wings within these domains. They exert their function, according to the 'selector gene' hypothesis, by regulating the expression of subordinate genes. Homoeotic genes also control their own expression and the expression of each other. The proteins encoded by these genes contain a domain, called a homoeodomain, that is strongly conserved, and that shows homologies to proteins that bind DNA and regulate transcription. Homoeoproteins have been shown to bind specific DNA sequences. We show here that the Drosophila homoeotic genes Ultrabithorax (Ubx) and Abdominal-B (Abd-B) code for proteins that are capable of activating transcription of reporter genes linked to specific cis-regulatory target sequences in transfected mammalian cells. Their activity, as well as their target specificity, is similar to that of a mammalian lymphoid-specific octamer transcription factor, OTF-2, which was recently found to contain a homoeodomain.

Animals↗
PubMed1988

A male accessory gland peptide that regulates reproductive behavior of female D. melanogaster.

The adult male accessory glands of D. melanogaster synthesize and secrete a peptide that represses female sexual receptivity and stimulates oviposition. Normally, this peptide is transferred to females during copulation; however, the peptide shows the same biological activity after purification and subsequent injection into the abdominal cavity of female virgins. Amino acid sequencing of the purified peptide and oligonucleotide-directed cDNA cloning established that the peptide consists of 36 amino acids. It appears to be synthesized as a precursor with a hydrophobic signal sequence of 19 residues at its N-terminal end. The precursor peptide is encoded by a short mRNA that accumulates exclusively in the male accessory gland. The gene has been localized by in situ hybridization to polytene chromosomes at 70A.

Amino Acid Sequence↗
PubMed1988

Domain of Ultrabithorax expression in Drosophila visceral mesoderm from autoregulation and exclusion.

Domains of differential homeotic gene activity are formed at specific positions along the anteroposterior axis of the early Drosophila embryo. Homeotic genes are required continuously throughout development, so that homeotic gene activity has to be maintained independently of the positional information provided in the early embryo. In the ectoderm, the domains of homeotic gene activity partially overlap, but we have found that in the visceral mesoderm at least three of these genes are expressed in adjacent and mutually exclusive domains. It has been proposed that stable, sharply demarcated domains of this type could be established if a homeotic gene product stimulated its own expression locally and inhibited the expression of other homeotic genes, which Meinhardt has termed autocatalysis and mutual exclusion respectively. Furthermore, autocatalysis of this kind can in principle account for the maintenance of homeotic gene activity throughout development. We find that the unique domain of Ultrabithorax (Ubx) expression in the visceral mesoderm is dependent both on autocatalysis and on an exclusion mechanism: Ubx product is required for its own synthesis, whereas the product of the posteriorly adjacent gene abdominal-A represses Ubx expression.

Animals↗
PubMed1988

Differential regulation of Ultrabithorax in two germ layers of Drosophila.

The homeotic gene Ultrabithorax (Ubx) is expressed in specific parts of Drosophila embryos: in a single metamer in the visceral mesoderm and forming a complex pattern limited to a broad domain in the ectoderm and in the somatic mesoderm. Here we use a linked beta-galactosidase gene to identify cis-acting regulatory sequences. In the visceral mesoderm, correct expression of Ubx depends on localized upstream sequences. In the ectoderm, all galactosidase-positive transformants show the same characteristic pattern. The repeated elements of this basal pattern appear to be a sub-pattern of engrailed (en) expression; they depend on en function as well as on sequences in the Ubx RNA leader. We use a mutant (Haltere-mimic) to show that sequences that normally restrict segmental expression of Ubx in the ectoderm are located downstream from the RNA leader.

Animals↗
PubMed1988

Evidence that the Abdominal-B r element function is conferred by a trans-regulatory homeoprotein.

The Abdominal-B gene is a homeotic gene located in the distal-most region of the bithorax complex (BX-C). Based on complementation analysis it has been proposed that the gene contains two separable genetic elements, called the m and r elements. The r element has a chiefly regulatory function confined to parasegment 14. In a reverse Northern screen of the distal-most BX-C DNA, we found four distinct areas that are transcribed in the embryo. One of the transcripts spans a large genomic region which, upon disruption by rearrangement breakpoints, causes loss of r element function. This transcript is expressed in the early embryo in a single domain apparently corresponding to parasegment 14. We propose that the small homeoprotein encoded by the transcript acts as a trans-regulator to confer r element function.

Amino Acid Sequence↗
PubMed1986

A CCAAT box confers cell-type-specific regulation on the Xenopus hsp70 gene in oocytes.

The Xenopus hsp70 gene is transiently heat-inducible in somatic cells; in injected oocytes, it is efficiently expressed without heat shock. I show here that this cell-type-specific and gene-specific activation is dependent on the presence of a heat shock regulatory element (HSE) and a CCAAT box in the hsp70 promoter; neither element by itself is sufficient for full activation. The same promoter elements are also required for heat inducibility in somatic cells. I propose that CAAT-binding transcription factor (CTF) interacts with heat shock transcription factor (HSTF), thereby enhancing the affinity of HSTF for the promoter; cell-type specificity could be explained by assuming either oocyte-specific forms or oocyte-specific high levels of CTF and/or HSTF.

Animals↗
PubMed1986

Heat shock regulatory elements function as an inducible enhancer in the Xenopus hsp70 gene and when linked to a heterologous promoter.

The Xenopus hsp70 promoter contains three copies of the consensus heat shock element (HSE) between positions -260 and -100. When the gene is transfected into mammalian cells, maximal heat-induced expression requires two HSEs in addition to a CCAAT box located next to the TATA box. The HSE-containing region can be separated from the CCAAT/TATA region without affecting expression of the gene, and it can enhance transcription of a linked beta-globin gene upon heat shock. It thus has the properties of a heat-inducible enhancer. Such an enhancer can also be generated by duplication of HSE sequences from the Drosophila hsp70 promoter, which were previously identified as an upstream promoter element and are known to bind a purified heat shock transcription factor in vitro.

Animals↗
PubMed1984

Xenopus hsp 70 genes are constitutively expressed in injected oocytes.

Xenopus heat-shock genes are transiently heat-inducible in somatic cells, but they are also subject to a long-term developmental control in oogenesis and early embryogenesis. In order to understand whether different genes or different promoter elements are involved in the two types of control, several genomic clones coding for Xenopus heat-shock proteins, hsp 70 and hsp 30, were isolated, characterised and tested for expression in oocytes and COS cells. Three isolated hsp 70 genes are nearly identical in their promoter and mRNA leader sequences, indicating that there is only one type of hsp 70 gene. These promoters contain a consensus sequence element (CT-GAA--TTC-AG) upstream of the TATA-box, which is presumably required for their transient heat-inducibility. The two isolated hsp 30 genes show 5'-flanking sequences similar to each other, except that one of them shows a homology disruption precisely around the consensus sequence element. The same gene contains a frameshift mutation in the protein coding part and, since it cannot be expressed after introduction into oocytes or COS cells, it is probably a pseudogene. The other hsp 30 gene is strongly heat-inducible in injected oocytes or transfected COS cells. In contrast, the hsp 70 genes are strongly heat-inducible in COS cells, but their expression is highly efficient in injected oocytes at the normal temperature and is not increased during heat shock. This represents correct cell type-specific regulation of a cloned reintroduced gene, since the endogenous hsp 70 genes are constitutively activated during oogenesis, leading to the accumulation of stored hsp 70 mRNA in oocytes.

Animals↗
PubMed1984

Developmental control of the heat shock response in Xenopus.

Xenopus cells express two major proteins on heat shock, designated hsp 70 and hsp 30. Several cDNA clones for the corresponding mRNAs were identified and sequenced. Inducibility and abundance of heat shock mRNAs in various cell types and developmental stages was determined by nuclease S1-mapping. The only cells found to contain hsp 70 mRNA without heat shock are the oocytes. The level of this stored hsp 70 mRNA is not increased by heat shock. After fertilization, hsp 70 mRNA becomes undetectable; it appears as a heat-inducible mRNA for the first time at gastrulation. After this stage, all somatic cell types accumulate hsp 70 mRNA to similar levels on heat shock, presumably by transcriptional activation of the hsp 70 genes. In contrast, hsp 30 mRNA is not detectable, even after heat shock, in oocytes or embryos that induce hsp 70 mRNA to high levels. Heat inducibility appears late in development--at the tadpole stage. However, the level of induced mRNA varies considerably in different adult tissues. This indicates that the Xenopus heat shock genes are not coordinately controlled. A long-term developmental control appears to be superimposed on the temporary heat inducibility of the heat shock genes: stage- or cell-type-specific conditions can lead to constitutive or repressed heat shock genes.

Aging↗
PubMed1983

A human tRNAGlu gene of high transcriptional activity.

A mixture of low molecular weight RNAs, in which only tRNAs were radiolabelled, was used as a hybridisation probe to select for tRNA-like sequences within a bank of human genomic DNA in lambda Charon 4A. A restriction enzyme digest of one of the selected lambda Charon 4A recombinants contained two fragments (2.4 Kb & 1.8 Kb) which hybridised tRNA and which, when subcloned into pAT153, were transcribed in Xenopus oocyte nuclei. Analysis of the subcloned 2.4 Kb fragment, which was of remarkably high transcriptional activity, revealed the presence of a single gene for tRNAGlu in the middle of the fragment. The sequence immediately preceding the gene has the potential for forming a tRNA-like structure.

Base Sequence↗
PubMed1982

A synthetic heat-shock promoter element confers heat-inducibility on the herpes simplex virus thymidine kinase gene.

Previous deletion analysis of the Drosophila hsp70 heat-shock promoter has identified a sequence upstream of the TATA box that is required for heat induction. This region contains homology to other heat-shock promoters, and it was proposed that the common sequence is an important element in the regulation of the heat-shock genes. We have constructed sequences similar to the consensus CT-GAA-TTC-AG from synthetic oligonucleotides and placed them upstream of the TATA box of the herpes virus thymidine kinase gene, in place of the normal upstream promoter element. The resultant genes are heat-inducible both in monkey COS cells and in Xenopus oocytes. We conclude that the transcriptional heat-shock response is mediated by some factor that interacts with this sequence.

Animals↗
PubMed1982

Expression of a Drosophila heat-shock protein in Xenopus oocytes: conserved and divergent regulatory signals.

On injection of cloned Drosophila hsp70 heat-shock genes into Xenopus oocytes, heat-inducible expression is observed: the level of correctly initiated transcripts is increased 20- to 100-fold on heat shock at 34 degrees C. We show that this induction is due to activation of the heat-shock gene promoter, and that the DNA sequences required for induction lie between 10 and 66 bases upstream from the transcription start site. Most heat-induced transcripts have a correct 3' end, and hsp70 mRNA activity is detectable after extraction of the RNA from oocytes and subsequent in vitro translation. Drosophila heat-shock protein (hsp70) is synthesised in the injected oocytes after heat-shock, but only at low temperature. Under heat-shock conditions, Drosophila hsp70 mRNA translation is reduced 10-fold as is translation of the normal (25 degrees C) mRNAs, whereas translation of the endogenous Xenopus hsp70 mRNA is strongly induced. Translation of Drosophila heat-shock mRNAs extracted from flies and injected into the oocytes is also reduced by heat-shock. This suggests that Xenopus oocytes do not recognise the translational regulatory signals of Drosophila heat-shock mRNAs. In contrast, the signals for heat-induced transcription must be strongly conserved between Xenopus and Drosophila.

Animals↗
PubMed1982

The heat-shock response in Xenopus oocytes is controlled at the translational level.

Xenopus laevis oocytes respond to high temperature (greater than 31 degrees C) by the synthesis of one major (70 kilodalton) protein and by a gradual reduction in the rate of normal protein synthesis. In contrast with most other cells, the heat-shock response of Xenopus oocytes is controlled exclusively at the translational level. Enucleated or alpha-amanitin-injected oocytes synthesize normal levels of heat-shock protein. Thus high temperature induces the translation of preformed heat-shock mRNA. This continues for more than a day after a shift back to a normal temperature, but ceases within 2 days. Heat-shock protein synthesis can be sequentially induced and inactivated in the same oocyte over several days. We conclude that an oocyte contains 10-100 pg of heat-shock mRNA, which is synthesized during oogenesis at the normal temperature, and which is stored in an inactive state by a "masking" mechanism.

Animals↗
PubMed1982

Mutations affecting the indirect flight muscles of Drosophila melanogaster.

The development of the indirect flight muscles of Drosophila melanogaster was studied by analysing mutations that cause flightlessness. Twenty-five mutations on the X-chromosome and two on the third chromosome were examined. The X-chromosomal mutations form ten complementation units. The ten loci were assigned preliminary map positions by meiotic recombination and deficiencies and duplications. The two autosomal mutations represent two genes. Gynandromorph analyses suggest that many of these mutations have their primary effect in the presumptive thoracic muscle region of the embryo. The mutations cause a variety of characteristic defects, such as absence of the bulk of the thoracic muscle mass, or absence of only one of the two fibrillar muscle groups. Electronmicroscopic studies of sixteen mutants representing all twelve loci reveal abnormal myofibrillar organization in some of these mutants, e.g. aberrant or missing Z-bands, or absence of the thin filaments. Mutant protein patterns, obtained by SDS-polyacrylamide gel electrophoresis, show the following differences from wild type: ten mutants are characterized by absence of reduction of the 54 K protein, and most mutants exhibit a reduction and modification of the 80 and 90 K proteins. The absence of reduction of the 54 K protein was strongly correlated with aberrant Z-bands.

Animals↗
PubMed1981

Usage of the three termination codons in a single eukaryotic cell, the Xenopus laevis oocyte.

Oocytes from Xenopus laevis were injected with purified amber (UAG), ochre (UAA), and opal (UGA) suppressor tRNAs from yeasts. The radioactively labeled proteins translated from the endogenous mRNAs were then separated on two-dimensional gels. All three termination codons are used in a single cell, the Xenopus laevis oocyte. But a surprisingly low number of readthrough polypeptides were observed from the 600 mRNAs studied in comparison to uninjected oocytes. The experimental data are compared with the conclusions obtained from the compilation of all available termination sequences on eukaryotic and prokaryotic mRNAs. This comparison indicates that the apparent resistance of natural termination codons against readthrough, as observed by the microinjection experiments, cannot be explained by tandem or very close second stop codons. Instead it suggests that specific context sequences around the termination codons may play a role in the efficiency of translation termination.

Animals↗
PubMed1981

The genes coding for tRNA Tyr of Drosophila melanogaster: localization of determination of the gene numbers.

Transfer RNA(Tyr) (anticodon G psi A) was isolated from Drosophila melanogaster by means of Sepharose 4B, RPC-5, and polyacrylamide gel electrophoresis. The rRNA was iodinated in vitro with Na125 I and hybridized in situ to salivary gland chromosomes from Drosophila. The genes of rRNA(Tyr) were localized in eight regions of the genome by autoradiography. Restriction enzyme analysis of genomic DNA indicated that the haploid Drosophila genome codes for about 23 tRNA(Tyr) genes. The regions 22F and 85A each contain four to five tRNA(Tyr) genes, whereas the regions 28C, 41AB, 42A, 42E, and 56D each contain two to three tRNA(Tyr) genes.

Animals↗