Autosomal P[ovoD1] dominant female-sterile insertions in Drosophila and their use in generating germ-line chimeras

Development ◽  
1993 ◽  
Vol 119 (4) ◽  
pp. 1359-1369 ◽  
Author(s):  
T.B. Chou ◽  
E. Noll ◽  
N. Perrimon
Keyword(s):  
Tissue Specificity ◽  
Germ Line ◽  
Egg Laying ◽  
P Element ◽  
Follicle Cells ◽  
Gtpase Activating Protein ◽  
Lethal Mutations ◽  
Sterile Technique ◽  
Dominant Female ◽  
Polarity Determination

The ‘dominant female-sterile’ technique used to generate germ-line mosaics in Drosophila is a powerful tool to determine the tissue specificity (germ line versus somatic) of recessive female-sterile mutations as well as to analyze the maternal effect of recessive zygotic lethal mutations. This technique requires the availability of germ-line-dependent, dominant female-sterile (DFS) mutations that block egg laying but do not affect viability. To date only one X-linked mutation, ovoD1 has been isolated that completely fulfills these criteria. Thus the ‘DFS technique’ has been largely limited to the X-chromosome. To extend this technique to the autosomes, we have cloned the ovoD1 mutation into a P-element vector and recovered fully expressed P[ovoD1] insertions on each autosomal arm. We describe the generation of these P[ovoD1] strains as well as demonstrate their use in generating germ-line chimeras. Specifically, we show that the Gap1 gene, which encodes a Drosophila homologue of mammalian GTPase-activating protein, is required in somatic follicle cells for embryonic dorsoventral polarity determination.

scholarly journals Requirement for cell-proliferation control genes in Drosophila oogenesis.

Genetics ◽  
1991 ◽  
Vol 127 (3) ◽  
pp. 525-533
Author(s):  
J Szabad ◽  
V A Jursnich ◽  
P J Bryant
Keyword(s):  
Cell Proliferation ◽  
Germ Line ◽  
Follicle Cell ◽  
Mitotic Recombination ◽  
Follicle Cells ◽  
Egg Development ◽  
Proliferation Control ◽  
Dominant Female ◽  
Female Germ Line ◽  
Cell Proliferation Control

Abstract Genes that are required for cell proliferation control in Drosophila imaginal discs were tested for function in the female germ-line and follicle cells. Chimeras and mosaics were produced in which developing oocytes and nurse cells were mutant at one of five imaginal disc overgrowth loci (fat, lgd, lgl, c43 and dco) while the enveloping follicle cells were normal. The chimeras were produced by transplantation of pole cells and the mosaics were produced by X-ray-induced mitotic recombination using the dominant female-sterile technique. The results show that each of the genes tested plays an essential role in the development or function of the female germ line. The fat, lgl and c43 homozygous germ-line clones fail to produce eggs, indicating a germ-line requirement for the corresponding genes. Perdurance of the fat+ gene product in mitotic recombination clones allows the formation of a few infertile eggs from fat homozygous germ-line cells. The lgd homozygous germ-line clones give rise to a few eggs with abnormal chorionic appendages, a defect thought to result from defective cell communication between the mutant germ-line and the nonmutant follicle cells. One allele of dco (dcole88) prevents egg development when homozygous in the germ line, whereas the dco18 allele has no effect on germ-line development. Fs(2)Ugra, a recently described follicle cell-dependent dominant female-sterile mutation, allowed the analysis of egg primordia in which fat, lgd or lgl homozygous mutant follicle cells surrounded normal oocytes. The results show that the fat and lgd genes are not required for follicle cell functions, while absence of lgl function in follicles prevents egg development.(ABSTRACT TRUNCATED AT 250 WORDS)

scholarly journals Two independent cis-acting elements regulate the sex- and tissue-specific expression ofyp3inDrosophila melanogaster

1995 ◽  
Vol 66 (1) ◽  
pp. 9-17 ◽  
Author(s):  
Elaine Ronaldson ◽  
Mary Bownes
Keyword(s):  
Fat Body ◽  
Germ Line ◽  
Ovarian Follicle ◽  
Yolk Protein ◽  
P Element ◽  
Follicle Cells ◽  
Cell Type Specificity ◽  
Specific Expression ◽  
Cis Acting ◽  
Ideal System

SummaryInDrosophila, the threeyolk protein(yp) genes are transcribed in a sex-, tissue- and developmentally specific manner, providing an ideal system in which to investigate the factors involved in their regulation. The yolk proteins are synthesized in the fat body of adult females, and in the ovarian follicle cells surrounding the developing oocyte during stages 8–10 of oogenesis. We report here an analysis of theyolk protein 3(yp3) gene and its flanking sequences by means of P-element mediated germ-line transformation and demonstrate that a 747 bp promoter region is sufficient to direct sex-specific expression in the female fat body and both the temporal- and cell-type-specificity of expression during oogenesis. Two elements that independently governyp3transcription in these tissues have been separated and no other sequences in the upstream, downstream or coding regions have been identified that are autonomously involved inyp3expression.

scholarly journals Two types of pole cells are present in the Drosophila embryo, one with and one without splicing activity for the third P-element intron

Development ◽  
1993 ◽  
Vol 117 (3) ◽  
pp. 885-893 ◽  
Author(s):  
S. Kobayashi ◽  
T. Kitamura ◽  
H. Sasaki ◽  
M. Okada
Keyword(s):  
Germ Cells ◽  
Germ Line ◽  
Primordial Germ Cells ◽  
Egg Laying ◽  
P Element ◽  
Drosophila Embryo ◽  
The Third ◽  
Pole Cells ◽  
Almost All ◽  
Second Peak

In Drosophila, it has been postulated that the third intron of the P-element is spliced only in germ-line cells. To test whether this postulate is applicable to pole cells, the progenitor cells of germ line, we carried out a histochemical assay to detect the splicing activity in embryos. The splicing activity was detected in pole cells and primordial germ cells. The activity increased to reach a maximum at 5–6 hours AEL (after egg laying), then decreased to an undetectable level by 8–9 hours AEL. The splicing activity showed a small second peak at 12–15 hours AEL. It was rather unexpected that not all pole cells were capable of splicing the third intron. Almost all pole cells that had the splicing activity at 5–6 hours AEL penetrated the embryonic gonads and differentiated into primordial germ cells. Our findings suggest that pole cells are selected to penetrate the gonads while they are migrating from the proctodeal cavity to the gonads. Furthermore, these results suggest that the machinery to splice the P-element is active in some pole cells, and that this activity is used for processing transcripts of genes that play important roles in the differentiation of pole cells into primordial germ cells.

scholarly journals Genetic and developmental analysis of polytene section 17 of the X chromosome of Drosophila melanogaster.

Genetics ◽  
1992 ◽  
Vol 130 (3) ◽  
pp. 569-583
Author(s):  
D F Eberl ◽  
L A Perkins ◽  
M Engelstein ◽  
A J Hilliker ◽  
N Perrimon
Keyword(s):  
Drosophila Melanogaster ◽  
X Chromosome ◽  
Developmentally Regulated ◽  
Present Evidence ◽  
Lethal Mutations ◽  
Sterile Technique ◽  
Dominant Female ◽  
Complementation Groups ◽  
Developmental Analysis ◽  
Chromosome Bands

Abstract Polytene section 17 of the X chromosome of Drosophila melanogaster, previously known to contain six putative lethal complementation groups important in oogenesis and embryogenesis, has here been further characterized genetically and developmentally. We constructed fcl+Y, a duplication of this region, which allowed us to conduct mutagenesis screens specific for the region and to perform complementation analyses (previously not possible). We recovered 67 new lethal mutations which defined 15 complementation groups within Df(1)N19 which deletes most of polytene section 17. The zygotic lethal phenotypes of these and preexisting mutations within polytene section 17 were examined, and their maternal requirements were analysed in homozygous germline clones using the dominant female sterile technique. We present evidence that an additional gene, which produces two developmentally regulated transcripts, is located in this region and is involved in embryogenesis, although no mutations in this gene were identified. In this interval of 37 to 43 polytene chromosome bands we have defined 17 genes, 12 (71%) of which are of significance to oogenesis or embryogenesis.

Monitoring positional information during oogenesis in adult Drosophila

Development ◽  
1988 ◽  
Vol 104 (2) ◽  
pp. 245-253 ◽  
Author(s):  
L. Fasano ◽  
S. Kerridge
Keyword(s):  
Egg Production ◽  
Germ Line ◽  
Spatial Localization ◽  
Follicle Cell ◽  
Positional Information ◽  
P Element ◽  
Follicle Cells ◽  
New Genes ◽  
Beta Galactosidase ◽  
Adult Drosophila

About 184P[lac, ry+]A insertions (O'Kane & Gehring, 1987) have been incorporated into the genome via P element-mediated transformation. The temporal-spatial localization of beta-galactosidase, synthesized by these insertions during oogenesis, is described. 32% present control levels of endogenous beta-galactosidase expression and 68% show novel patterns. 13% of the insertions are germline-specific; 33%, follicle-cell-specific; 20% are expressed in both germ line and follicle cells; and 2%, specific to the germarium. Several lines exhibit strict temporal-spatial localizations of beta-galactosidase; notably those expressed in specific populations of follicle cells. The results are discussed with respect to some of the positional information encoded in the genome to which the insertions respond, the use of the insertions as markers for cell differentiation and the potential of the technique for isolating new genes involved in egg production.

scholarly journals The Autosomal FLP-DFS Technique for Generating Germline Mosaics in Drosophila melanogaster

Genetics ◽  
1996 ◽  
Vol 144 (4) ◽  
pp. 1673-1679 ◽  
Author(s):  
Tze-bin Chou ◽  
Norbert Perrimon
Keyword(s):  
Drosophila Melanogaster ◽  
Maternal Effect ◽  
Tissue Specificity ◽  
Efficient Production ◽  
Site Specific ◽  
Lethal Mutations ◽  
Recombination System ◽  
Dominant Female ◽  
Selection For ◽  
Female Germline

The production of female germline chimeras is invaluable for analyzing the tissue specificity of recessive female sterile mutations as well as detecting the maternal effect of recessive zygotic lethal mutations. Previously, we developed the “FLP-DFS” technique to efficiently generate germline clones. This technique uses the X-linked germline-dependent dominant female sterile mutation ovo  D1 as a selection for the detection of germline recombination events, and the FLP-FRT recombination system to promote site-specific chromosomal exchange. This method allows the efficient production of germline mosaics only on the X chromosome. In this paper we have built chromosomes that allow the use of this technique to the autosomes. We describe the various steps involved in the development of this technique as well as the properties of the chromosomes utilized.

scholarly journals Genetic and Molecular Characterization of sting, a Gene Involved in Crystal Formation and Meiotic Drive in the Male Germ Line of Drosophila melanogaster

Genetics ◽  
1999 ◽  
Vol 151 (2) ◽  
pp. 749-760 ◽  
Author(s):  
Armin Schmidt ◽  
Gioacchino Palumbo ◽  
Maria P Bozzetti ◽  
Patrizia Tritto ◽  
Sergio Pimpinelli ◽  
...  
Keyword(s):  
Amino Acids ◽  
Drosophila Melanogaster ◽  
Null Allele ◽  
Germ Line ◽  
Meiotic Drive ◽  
P Element ◽  
Crystal Formation ◽  
Male Sterile ◽  
Male Sterile Mutation

Abstract The sting mutation, caused by a P element inserted into polytene region 32D, was isolated by a screen for male sterile insertions in Drosophila melanogaster. This sterility is correlated with the presence of crystals in spermatocytes and spermatids that are structurally indistinguishable from those produced in males carrying a deficiency of the Y-linked crystal (cry) locus. In addition, their morphology is needle-like in Ste+ flies and star-shaped in Ste flies, once again as observed in cry– males. The sti mutation leads to meiotic drive of the sex chromosomes, and the strength of the phenomenon is correlated with the copy number of the repetitive Ste locus. The same correlation is also true for the penetrance of the male sterile mutation. A presumptive sti null allele results in male sterility and lethal maternal effect. The gene was cloned and shown to code for a putative protein that is 866 amino acids long. A C-terminal domain of 82 amino acids is identified that is well conserved in proteins from different organisms. The gene is expressed only in the germline of both sexes. The interaction of sting with the Ste locus can also be demonstrated at the molecular level. While an unprocessed 8-kb Ste primary transcript is expressed in wild-type males, in X/Y homozygous sti males, as in X/Y cry– males, a 0.7-kb mRNA is produced.

pitkinD, a Novel Gain-of-Function Enhancer of Position-Effect Variegation, Affects Chromatin Regulation During Oogenesis and Early Embryogenesis in Drosophila

Genetics ◽  
2001 ◽  
Vol 157 (3) ◽  
pp. 1227-1244 ◽  
Author(s):  
Steffi Kuhfittig ◽  
János Szabad ◽  
Gunnar Schotta ◽  
Jan Hoffmann ◽  
Endre Máthé ◽  
...  
Keyword(s):  
Germ Line ◽  
Position Effect ◽  
Early Embryogenesis ◽  
Position Effect Variegation ◽  
Chromatin Regulation ◽  
Gain Of Function ◽  
Position Effects ◽  
Dominant Female ◽  
Effect Variegation ◽  
Female Germ Line

Abstract The vast majority of the >100 modifier genes of position-effect variegation (PEV) in Drosophila have been identified genetically as haplo-insufficient loci. Here, we describe pitkinDominant (ptnD), a gain-of-function enhancer mutation of PEV. Its exceptionally strong enhancer effect is evident as elevated spreading of heterochromatin-induced gene silencing along euchromatic regions in variegating rearrangements. The ptnD mutation causes ectopic binding of the SU(VAR)3-9 heterochromatin protein at many euchromatic sites and, unlike other modifiers of PEV, it also affects stable position effects. Specifically, it induces silencing of white+ transgenes inserted at a wide variety of euchromatic sites. ptnD is associated with dominant female sterility. +/+ embryos produced by ptnD/+ females mated with wild-type males die at the end of embryogenesis, whereas the ptnD/+ sibling embryos arrest development at cleavage cycle 1-3, due to a combined effect of maternally provided mutant product and an early zygotic lethal effect of ptnD. This is the earliest zygotic effect of a mutation so far reported in Drosophila. Germ-line mosaics show that ptn+ function is required for normal development in the female germ line. These results, together with effects on PEV and white+ transgenes, are consistent with the hypothesis that the ptn gene plays an important role in chromatin regulation during development of the female germ line and in early embryogenesis.

Identification of Chromosome Inheritance Modifiers in Drosophila melanogaster

Genetics ◽  
2001 ◽  
Vol 157 (4) ◽  
pp. 1623-1637 ◽  
Author(s):  
Kenneth W Dobie ◽  
Cameron D Kennedy ◽  
Vivienne M Velasco ◽  
Tory L McGrath ◽  
Juliani Weko ◽  
...  
Keyword(s):  
Drosophila Melanogaster ◽  
Biological Activity ◽  
Cancer Progression ◽  
Birth Defects ◽  
Mitotic Chromosome ◽  
P Element ◽  
Inverse Pcr ◽  
Gtpase Activating Protein ◽  
New Genes ◽  
Genomic Analyses

Abstract Faithful chromosome inheritance is a fundamental biological activity and errors contribute to birth defects and cancer progression. We have performed a P-element screen in Drosophila melanogaster with the aim of identifying novel candidate genes involved in inheritance. We used a “sensitized” minichromosome substrate (J21A) to screen ∼3,000 new P-element lines for dominant effects on chromosome inheritance and recovered 78 Sensitized chromosome inheritance modifiers (Scim). Of these, 69 decreased minichromosome inheritance while 9 increased minichromosome inheritance. Fourteen mutations are lethal or semilethal when homozygous and all exhibit dramatic mitotic defects. Inverse PCR combined with genomic analyses identified P insertions within or close to genes with previously described inheritance functions, including wings apart-like (wapl), centrosomin (cnn), and pavarotti (pav). Further, lethal insertions in replication factor complex 4 (rfc4) and GTPase-activating protein 1 (Gap1) exhibit specific mitotic chromosome defects, discovering previously unknown roles for these proteins in chromosome inheritance. The majority of the lines represent mutations in previously uncharacterized loci, many of which have human homologs, and we anticipate that this collection will provide a rich source of mutations in new genes required for chromosome inheritance in metazoans.

logjam Encodes a Predicted EMP24/GP25 Protein That Is Required for Drosophila Oviposition Behavior

Genetics ◽  
2003 ◽  
Vol 164 (1) ◽  
pp. 173-186
Author(s):  
Ginger E Carney ◽  
Barbara J Taylor
Keyword(s):  
Intracellular Trafficking ◽  
Oviposition Behavior ◽  
Egg Laying ◽  
P Element ◽  
Female Reproduction ◽  
Multiple Functions ◽  
Cytoplasmic Vesicles ◽  
Egg Chambers ◽  
Low Levels ◽  
Overlapping Transcripts

Abstract A newly characterized Drosophila melanogaster gene, logjam (loj), functions in female reproduction by modulating oviposition behavior. The locus encodes at least six overlapping transcripts with unique 5′ ends. P-element mutants that express very low levels of loj transcripts are unable to oviposit mature eggs. This phenotype can be rescued by the introduction of a transgene expressing the most abundant loj transcript. As for many genes that specify behavioral outputs, loj is present in the adult central nervous system (CNS). Interestingly, it is also observed in vitellogenic egg chambers, suggesting that there may be multiple functions for this gene in egg-laying behavior. loj encodes a predicted protein with homology to the EMP24/GP25 transmembrane components of cytoplasmic vesicles and likely functions in intracellular trafficking.

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