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J A Lepesant

Publications and source records attributed to J A Lepesant.

11 recordsLinked to original sources
PubMed1992

Spatial distribution of the Sm antigen in Drosophila early embryos.

Anti-Sm antibodies recognize the major small nuclear RNA-protein particles (snRNPs) involved in pre-mRNA processing. The spatial distribution of the snRNPs has been investigated in Drosophila embryos up to the cellularization stage (cycle 14), using the Y12 anti-Sm antibody. Our results show that: 1) all or most of the Sm antigen is localized in the cytoplasm of the syncytial blastoderm until the 12th cycle of division, in both the nuclear and cytoplasmic compartments at cycle 13, and then in the nuclei at cycle 14 and later. This relocalization takes place when zygotic transcriptional activation occurs; 2) at the subcellular level, the Sm antigen localizes in a speckled pattern and in foci-like structures within the nucleus of Drosophila blastoderm embryos; 3) strikingly, some nuclei of embryos at the 14th cycle appear to contain more snRNPs than others. The position of these nuclei differs from one embryo to another, and their distribution does not resemble any known developmental pattern of Drosophila embryogenesis. We propose that random differences in snRNP concentration may serve as an epigenetic signal for stochastic events occurring during development.

Animals↗
PubMed1992

The Drosophila pourquoi-pas?/wings-down zinc finger protein: oocyte nucleus localization and embryonic requirement.

The pourquoi-pas? (pqp) gene of Drosophila melanogaster encodes a Cys2/His2 zinc finger protein and is abundantly transcribed in adult ovaries. During oogenesis, we immunodetected the pqp protein in the nucleus of nurse cells at stages 1-6, in a spherical structure within the oocyte nucleus at stages 7-9, and uniformly distributed in the oocyte nucleus and in nurse cell nuclei at later stages. The pqp protein is also present at a lower level in the nuclei of follicle cells, embryos, and larvae. By means of a polymerase chain reaction (PCR) screen, we recovered three independent and phenotypeless P-element insertions at the pqp locus. In a second step, two excision-induced deletions of the pqp gene were isolated after mobilization of one of these P elements. The pqp mutants display zygotic (spread and drooping wings, cross-vein defects, extra bristles) and maternal (embryonic lethality) recessive phenotypes. The chromosomal position (98EF) of the pqp gene and the drooping wing phenotype of the pqp mutants agree with the hypothesis that the pqp gene is the wings down (wdn) gene for which T.H. Morgan isolated (and lost) mutants in the 1920s. This is the first reported occurrence of a zinc finger protein in the nucleus of the Drosophila oocyte.

Animals↗
PubMed1991

Drosophila fat body protein P6 and alcohol dehydrogenase are derived from a common ancestral protein.

Drosophila melanogaster alcohol dehydrogenase is an example of convergent evolution: it is not related to the ADHs of other organisms, but to short-chain dehydrogenases, which until now have been found only in bacteria and in mammalian steroid hormone metabolism. We present evidence that the Drosophila ADH is phylogenetically more closely related to P6, another highly expressed protein from the fat body of Drosophila, than it is to the short-chain dehydrogenases. The polypeptide sequence of P6 was inferred from DNA sequence analysis. Both ADH and P6 polypeptides have retained a high structural similarity with respect to the Chou-Fasman prediction of secondary structure and hydropathy. P6 is also homologous to the 25-kd protein from the fat body of Sarcophaga peregrina, whose sequence we have reexamined. The evolution of the P6-ADH family of proteins is characterized by a dramatic increase in the methionine content of P6. Methionine accounts for 20% of P6 amino acids. This is in contrast with the absence of this amino acid in mature ADH. There is evidence that P6 and the 25-kd protein have undergone a parallel and independent enrichment in methionine. When corrected for this, the rate of amino acid replacement shows that the P6-25-kd lineage diverged from insect ADH shortly before the divergence of the ADH gene (Adh) from its 3'-duplication (Adh-dup).

Alcohol Dehydrogenase↗
PubMed1991

Genetic analysis of the cellularization of the Drosophila embryo.

The synchronous cellularization of the Drosophila embryo at the blastoderm stage provides a unique system for studying the molecular mechanisms involved in cytokinesis, using genetical and biochemical approaches. The cellularization process requires the major components of the embryonic cytoskeleton that are deposited into the egg during oogenesis. Genetical analysis indicates that it requires also the products of additional maternally-acting genes, as well as that of a limited set of zygotically-acting genes. The cellularization defective phenotypes associated with small deficiencies uncovering these latter loci reveal specific steps within this complex process. The molecular analysis of these genes will ultimately provide meaningful insights into the normal process of cellularization. Among them, the serendipity alpha gene encodes a membrane-associated protein, which is exclusively accumulated during cellularization, and is required for the reorganization of the microfilaments as the onset of cellularization.

Actins↗
PubMed1990

Structure of the ecdysone-inducible P1 gene of Drosophila melanogaster.

The P1 gene codes for a major RNA, which accumulates specifically in the fat body cells at the late third larval stage of Drosophila melanogaster development under the positive control of the insect molting hormone 20-hydroxyecdysone. The primary structure of the P1 gene and the 5' upstream flanking region to position -776 relative to the transcription start was determined by sequence analysis of a cloned genomic DNA segment and two cDNAs containing sequences complementary to the 5' and 3' ends of the P1 transcript. The RNA coding region spans 3469 nucleotides and contains a 59-base-pair intron close to its 5' end, as predicted by computer analysis and established by S1 nuclease protection, primer extension and cDNA sequencing. The predicted P1 polypeptide contains 1030 amino acids, including a putative 16-amino acid signal peptide and two stretches of 12 and 11 aspartic and asparagine residues. Short stretches of nucleotide sequences similar to sequences located in the 5' regions of other genes expressed in the D. melanogaster fat body were found in the proximal promoter and transcribed region of the P1 gene.

Amino Acid Sequence↗
PubMed1990

[Homeotic genes].

Homeotic genes occupy a central position in the hierarchy of genes controlling the early steps of embryonic development in Drosophila. They are involved in the specification of the individual identity of each segment of the insect's body. Most homeotic genes of Drosophila are located in two large gene clusters, the Antennapedia complex (ANT-C) and the Bithorax complex (BX-C). These genes are expressed in two partially overlapping domains whose positions along the anterior-posterior axis of the organism are colinear with their position within each complex. Five genes in the ANT-C are involved in the specification of the identity of some segments of the head and of that of the first and second thoracic segments. The three genes of the BX-C determine the identities of the posterior compartment of the second and third thoracic segments and of the eight abdominal segments. Molecular studies have revealed that the proteins coded by the homeotic genes share a 60 amino acid motif, the homeo box, whose helix-turn-helix structure enables them to bind as transcription factors to specific DNA sequences in the cis-acting regulatory regions of their target genes. The high degree of phylogenetic conservation of the homeobox has made possible the identification and the isolation of more than fifty homeobox genes (Hox genes) in many eukaryotic organisms including nematode, Xenopus, mouse and man. In the mouse and human genomes Hox genes are clustered in tandem in four large gene complexes located on separate chromosomes. The relative positions of the genes within the clusters and the positions of the anterior boundaries of their expression domains along the anterior-posterior axis of the central nervous system and prevertebral column of the mouse embryo were found to be colinear. The similarities between the structure, organization and pattern of expression of the homeotic genes of Drosophila and of the homeobox genes of vertebrates suggest that some basic principles of embryonic organization and development have been preserved during the long period of evolution that has elapsed since the divergence of the phylogenetic lineages leading to vertebrates and arthropods.

Animals↗
PubMed1978

Ecdysone-inducible functions of larval fat bodies in Drosophila.

Late in the third instar larval stage of Drosophila melanogaster, the titer of the steroid hormone ecdysone increases sharply. This increase is blocked in the temperature-sensitive mutant ecd(1) after a temperature shift from 20 degrees C to 29 degrees C. The mutant was used to prepare three samples of late third instar larvae with different titers of ecdysone; the titer was low in one sample because of an earlier temperature shift, high in a second sample because the larvae were subsequently transferred to ecdysone-supplemented food, and also high in a third sample that was kept at 20 degrees C, providing a control for normal development. The effect of the high titer of ecdysone on proteins of the larval fat bodies was examined by comparing two-dimensional gel electrophoresis patterns of total proteins in stained gels. There were proteins at five positions in the gels for the high-ecdysone samples that were not detected at the corresponding positions in the gel for the low-ecdysone sample. The effect of ecdysone on these proteins was further studied by injecting [(35)S]methionine into the larvae at both early and late third instar stages, in order to label proteins synthesized before and after the increase in ecdysone titer. The results indicate that ecdysone induces two major responses in the fat bodies; certain proteins that were synthesized earlier in the fat bodies and secreted into the hemolymph are incorporated back into the fat bodies, and other proteins are newly synthesized. Attempts to induce prematurely the synthesis of the new proteins by exposing early third instar larvae to exogenous ecdysone were unsuccessful, suggesting that development must proceed further before the fat bodies can respond to ecdysone. By in vitro translation of RNA isolated from fat bodies of low-and high-ecdysone samples of larvae, it was shown that ecdysone greatly increases the amount of translatable messenger RNA for one of the newly synthesized proteins. A clone of DNA complementary to the induced messenger RNA has been isolated from a population of lambda bacteriophage carrying segments of the Drosophila genome. Using the cloned DNA to measure amounts of complementary poly(A)-RNA in the fat bodies by DNA.RNA hybridization, we detected about 50 times more complementary poly(A)-RNA in the high-ecdysone sample of larvae than in the low-ecdysone sample. This finding provides direct evidence that ecdysone induces an increase in the amount of the messenger RNA. The ecdysone-induced appearance of a major messenger RNA in late third instar larval fat bodies represents a developmental response to ecdysone that appears to be gene-specific, tissue-specific, and stage-specific, and it has exceptionally favorable features for further molecular studies of the control of gene expression by a steroid hormone.

Adipose Tissue↗
PubMed1977

Presence of a third sucrose hydrolyzing enzyme in Bacillus subtilis: constitutive levanase synthesis by mutants of Bacillus subtilis Marburg 168.

A beta-D-fructofuranosidase -- called levanase -- capable of the hydrolysis of sucrose, inulin and levans has been identified in Bacillus subtilis Marburg. This enzyme can not be detected in strain 168. However, sacL mutations -- mapped on the chromosome of strain 168 between the pheA and aroD reference markers -- lead to constitutive levanase synthesis. This synthesis is repressed by carbon sources such as glucose, glycerol or sucrose.

Bacillus subtilis↗
PubMed1975

Revision of the linkage map of Bacillus subtilis 168: indications for circularity of the chromosome.

A revision of the linkage map of the Bacillus subtilis 168 chromosome has been undertaken with the use of the generalized transducing phage PBS1. The mapping of four new markers (narB1, mtlB1, aroI906, and tre-12) has allowed a determination of the relative orientation of the purB-dal segment and its linkage with the lin markers. The chromosomal segment comprised between the sacQ36 and gtaA12 markers has been linked with the narA1, ctrA1, and sacA321 markers. The recA1 marker has been mapped relative to the thyA and citB17 markers. Indications of linkage have been found between the tre-12 and catA markers and the aroG932 and sacQ36 markers. According to these results, a circular genetic map of the chromosome of B. subtilis 168 is presented. Taken together, the transduction data and the order of marker replication determined by Harford in the accompanying paper support strongly the hypothesis of a symmetrical and fully bidirectional mode of replication for the B. subtilis 168 chromosome.

Bacillus subtilis↗