scholarly journals Molecular genetics of the Drosophila melanogaster ovo locus, a gene required for sex determination of germline cells.

Genetics ◽  
1992 ◽  
Vol 130 (4) ◽  
pp. 791-803
Author(s):  
M D Garfinkel ◽  
A R Lohe ◽  
A P Mahowald
Keyword(s):  
Drosophila Melanogaster ◽  
Sex Determination ◽  
Genomic Dna ◽  
Transcription Unit ◽  
Complementation Group ◽  
P Element ◽  
Female Sterility ◽  
Germline Cells ◽  
Female Specific ◽  
Female Germline

Abstract The Drosophila melanogaster ovo gene is required for survival and differentiation of female germline cells, apparently playing a role in germline sex determination. We recovered 60 kb of genomic DNA from its genetic location at 4E1,2 on the X chromosome. A transcription unit coding for an apparently female-specific germline-dependent 5-kb poly(A)+ RNA size class is located substantially in a 7-kb region, within which three DNA-detectable lesions for mutations that inactivate the ovo function are located at two sites approximately 4 kb apart. The breakpoint of a deficiency that removes the neighboring lethal complementation group shavenbaby (svb) but leaves the ovo function intact maps approximately 5 kb to the molecular left of the leftmost ovo mutant site. A class of mutations that inactivates both the svb function and the ovo function affects genomic DNA between the two ovo sites. Sequences required for the two genetic functions are partly overlapping. In spite of this overlap, P element-mediated gene transfer of a 10-kb genomic DNA segment containing the 5-kb poly(A)+ RNA transcription unit rescues the female sterility phenotypes of ovo mutations, but not the svb lethality.

Molecular cloning and genetic analysis of the suppressor-of-white-apricot locus from Drosophila melanogaster

1987 ◽  
Vol 7 (7) ◽  
pp. 2498-2505
Author(s):  
Z Zachar ◽  
D Garza ◽  
T B Chou ◽  
J Goland ◽  
P M Bingham
Keyword(s):  
Drosophila Melanogaster ◽  
Gene Transfer ◽  
Genetic Analysis ◽  
Developmental Stages ◽  
Chromosomal Region ◽  
Transcription Unit ◽  
Complementation Group ◽  
P Element ◽  
Allele Specific ◽  
New Mutations

We report genetic and molecular analysis of the suppressor-of-white-apricot [su(wa)] locus, one of several retrotransposon insertion allele-specific suppressor loci in Drosophila melanogaster. First, we isolated and characterized eight new mutations allelic to the original su(wa)1 mutation. These studies demonstrated that su(wa) mutations allelic to su(wa)1 affected a conventional D. melanogaster complementation group. Second, we cloned the chromosomal region containing the su(wa) complementation group by P element transposon tagging. The ca. 14-kilobase region surrounding the su(wa) complementation group contained five distinct transcription units, each with a different developmentally programmed pattern of expression. Third, we used a modified procedure for P-mediated gene transfer to identify the transcription unit corresponding to su(wa) by gene transfer. Fourth, we found that the presumptive su(wa) transcription unit produced a family of transcripts (ranging from ca. 3.5 to ca. 5.2 kilobases) in all developmental stages, tissue fractions, and cell lines we examined, suggesting that the gene is universally expressed.

scholarly journals Molecular cloning and genetic analysis of the suppressor-of-white-apricot locus from Drosophila melanogaster.

1987 ◽  
Vol 7 (7) ◽  
pp. 2498-2505 ◽  
Author(s):  
Z Zachar ◽  
D Garza ◽  
T B Chou ◽  
J Goland ◽  
P M Bingham
Keyword(s):  
Drosophila Melanogaster ◽  
Gene Transfer ◽  
Genetic Analysis ◽  
Developmental Stages ◽  
Chromosomal Region ◽  
Transcription Unit ◽  
Complementation Group ◽  
P Element ◽  
Allele Specific ◽  
New Mutations

We report genetic and molecular analysis of the suppressor-of-white-apricot [su(wa)] locus, one of several retrotransposon insertion allele-specific suppressor loci in Drosophila melanogaster. First, we isolated and characterized eight new mutations allelic to the original su(wa)1 mutation. These studies demonstrated that su(wa) mutations allelic to su(wa)1 affected a conventional D. melanogaster complementation group. Second, we cloned the chromosomal region containing the su(wa) complementation group by P element transposon tagging. The ca. 14-kilobase region surrounding the su(wa) complementation group contained five distinct transcription units, each with a different developmentally programmed pattern of expression. Third, we used a modified procedure for P-mediated gene transfer to identify the transcription unit corresponding to su(wa) by gene transfer. Fourth, we found that the presumptive su(wa) transcription unit produced a family of transcripts (ranging from ca. 3.5 to ca. 5.2 kilobases) in all developmental stages, tissue fractions, and cell lines we examined, suggesting that the gene is universally expressed.

scholarly journals A hobo Transgene That Encodes the P-Element Transposase in Drosophila melanogaster: Autoregulation and Cytotype Control of Transposase Activity

Genetics ◽  
2002 ◽  
Vol 161 (1) ◽  
pp. 195-204 ◽  
Author(s):  
Michael J Simmons ◽  
Kevin J Haley ◽  
Craig D Grimes ◽  
John D Raymond ◽  
Jarad B Niemi
Keyword(s):  
Drosophila Melanogaster ◽  
Gonadal Dysgenesis ◽  
P Element ◽  
Hsp70 Gene ◽  
Alternate Splicing ◽  
Male And Female ◽  
P Elements ◽  
Male Germline ◽  
Female Germline ◽  
Transposase Expression

Abstract Drosophila were genetically transformed with a hobo transgene that contains a terminally truncated but otherwise complete P element fused to the promoter from the Drosophila hsp70 gene. Insertions of this H(hsp/CP) transgene on either of the major autosomes produced the P transposase in both the male and female germlines, but not in the soma. Heat-shock treatments significantly increased transposase activity in the female germline; in the male germline, these treatments had little effect. The transposase activity of two insertions of the H(hsp/CP) transgene was not significantly greater than their separate activities, and one insertion of this transgene reduced the transposase activity of P(ry+, Δ2-3)99B, a stable P transgene, in the germline as well as in the soma. These observations suggest that, through alternate splicing, the H(hsp/CP) transgene produces a repressor that feeds back negatively to regulate transposase expression or function in both the somatic and germline tissues. The H(hsp/CP) transgenes are able to induce gonadal dysgenesis when the transposase they encode has P-element targets to attack. However, this ability and the ability to induce P-element excisions are repressed by the P cytotype, a chromosomal/cytoplasmic state that regulates P elements in the germline.

scholarly journals Cytoplasmic dynein (ddlc1) mutations cause morphogenetic defects and apoptotic cell death in Drosophila melanogaster.

1996 ◽  
Vol 16 (5) ◽  
pp. 1966-1977 ◽  
Author(s):  
T Dick ◽  
K Ray ◽  
H K Salz ◽  
W Chia
Keyword(s):  
Drosophila Melanogaster ◽  
Light Chain ◽  
Blot Analysis ◽  
Cytoplasmic Dynein ◽  
P Element ◽  
Female Sterility ◽  
Dynein Light Chain ◽  
Loss Of Function ◽  
Light Chain Gene

We report the molecular and genetic characterization of the cytoplasmic dynein light-chain gene, ddlc1, from Drosophila melanogaster. ddlc1 encodes the first cytoplasmic dynein light chain identified, and its genetic analysis represents the first in vivo characterization of cytoplasmic dynein function in higher eucaryotes. The ddlc1 gene maps to 4E1-2 and encodes an 89-amino-acid polypeptide with a high similarity to the axonemal 8-kDa outer-arm dynein light chain from Chlamydomonas flagella. Developmental Northern (RNA) blot analysis and ovary and embryo RNA in situ hybridizations indicate that the ddlc1 gene is expressed ubiquitously. Anti-DDLC1 antibody analyses show that the DDLC1 protein is localized in the cytoplasm. P-element-induced partial-loss-of-function mutations cause pleiotropic morphogenetic defects in bristle and wing development, as well as in oogenesis, and hence result in female sterility. The morphological abnormalities found in the ovaries are always associated with a loss of cellular shape and structure, as visualized by a disorganization of the actin cytoskeleton. Total-loss-of-function mutations cause lethality. A large proportion of mutant animals degenerate during embryogenesis, and the dying cells show morphological changes characteristic of apoptosis, namely, cell and nuclear condensation and fragmentation, as well as DNA degradation. Cloning of the human homolog of the ddlc1 gene, hdlc1, demonstrates that the dynein light-chain 1 is highly conserved in flies and humans. Northern blot analysis and epitope tagging show that the hdlc1 gene is ubiquitously expressed and that the human dynein light chain 1 is localized in the cytoplasm. hdlc1 maps to 14q24.

scholarly journals Genetic study of motor functions in Drosophila melanogaster

2012 ◽  
Vol 10 (1) ◽  
pp. 51-61 ◽  
Author(s):  
Sergey A Fedotov ◽  
Julia V Bragina ◽  
Nataliya G Besedina ◽  
Larisa V Danilenkova ◽  
Elena A Kamysheva ◽  
...  
Keyword(s):  
Drosophila Melanogaster ◽  
Motor Activity ◽  
Genomic Dna ◽  
Molecular Mechanisms ◽  
Central Pattern Generators ◽  
Courtship Song ◽  
P Element ◽  
Rhythmic Motor ◽  
Pattern Generators ◽  
First Time

To investigate molecular mechanisms of central pattern generators (CPG s) functioning, we carried out a screening of collection of Drosophila P-insertional mutants for strong deviations in locomotion and courtship song. In 21 mutants, the site of the P-insertion was localized by sequencing of the fragments of genomic DNA flanking the P-element. Bioinformational analysis revealed a list of candidate genes, potential players in development and functioning of CPG s. Possible involvement of certain identified genes in rhythmic motor activity is suggested for the first time (CG15630, Map205).

scholarly journals Construction, stable transformation, and function of an amber suppressor tRNA gene in Drosophila melanogaster

1989 ◽  
Vol 86 (17) ◽  
pp. 6696-6698 ◽  
Author(s):  
F A Laski ◽  
S Ganguly ◽  
P A Sharp ◽  
U L RajBhandary ◽  
G M Rubin
Keyword(s):  
Drosophila Melanogaster ◽  
Trna Gene ◽  
P Element ◽  
Female Sterility ◽  
Suppressor Trna ◽  
Amber Codon ◽  
Amber Suppressor ◽  
Chloramphenicol Acetyltransferase Gene ◽  
Male And Female Sterility ◽  
And Function

Drosophila melanogaster strains with a stably incorporated amber suppressor tRNA gene have been generated. A tRNATyr gene was site specifically mutated to produce an anticodon sequence that recognizes the amber codon and then introduced into Drosophila by using P-element-mediated transformation. Transformants from four integration events were recovered. Two integrations resulted in both male and female sterility, whereas the other two resulted in male sterility but female fertility. Strains derived from the two female-fertile integration events were shown to have a low level of amber-suppressing activity by their ability to suppress an amber mutation in a chloramphenicol acetyltransferase gene.

scholarly journals P-element-induced control mutations at the r gene of Drosophila melanogaster.

1985 ◽  
Vol 5 (10) ◽  
pp. 2567-2574 ◽  
Author(s):  
S Tsubota ◽  
M Ashburner ◽  
P Schedl
Keyword(s):  
Drosophila Melanogaster ◽  
The Other ◽  
P Element ◽  
Female Sterility ◽  
R Gene ◽  
Temporal Expression ◽  
Base Pairs ◽  
P Elements ◽  
Adult Females ◽  
Regulatory Mutations

The P-M hybrid dysgenesis system was used to produce five putative regulatory mutations at the rudimentary locus, r. All five mutations were the result of insertions at the 5' end of the gene, upstream of the proposed start of transcription. All of the mutants displayed a leaky wing phenotype, and four of the mutants showed an uncoupling of the wing and female-sterility phenotypes, suggesting that they altered the normal spatial and temporal expression of the r gene. Four of the insertions were P elements. The fifth insertion, which was larger than an intact P element, consisted of a small P element connected to non-P-element DNA. Two of the mutants produced very little r transcript in adult females and were clustered 80 to 150 base pairs upstream of the start of transcription. The other three mutants had higher levels of r transcript in adult females and were clustered 440 to 500 base pairs upstream of the start of transcription. All of the data suggest that the insertions are in a 5' noncoding region of the r gene involved in the control of its spatial and temporal expression.

scholarly journals Isolation and characterization of the prune locus of Drosophila melanogaster.

Genetics ◽  
1991 ◽  
Vol 128 (2) ◽  
pp. 373-380
Author(s):  
D H Teng ◽  
L B Bender ◽  
C M Engele ◽  
S Tsubota ◽  
T Venkatesh
Keyword(s):  
Drosophila Melanogaster ◽  
Transcription Unit ◽  
P Element ◽  
Isolation And Characterization ◽  
Body Tissues ◽  
Length Polymorphisms ◽  
Pigment Biosynthesis ◽  
Adult Head

Abstract The complementary lethal interaction between the prune (pn) and Killer of prune loci of Drosophila melanogaster is an unusual and highly specific phenomenon. A lesion in pn results in a brownish-purple color of the compound eyes, while the conditional dominant Killer of prune mutation exhibits no phenotype by itself. However, a hemizygous or homozygous pn mutant carrying a copy of the Killer of prune gene dies during the late second to third instar stage of larval development. As a step toward understanding the molecular nature of this lethality and the role of pn in pigment biosynthesis, we have cloned the pn locus by using a transposon tag in the P element-induced allele, pn38. In addition, seven independent revertant lines were generated by the remobilization of transposons in pn38. The pn gene is located in a region that is transcriptionally active, and the isolated cDNAs that correspond to this area fall into three transcription units: I, II and III. Southern analysis shows that the restriction fragment length polymorphisms in five pn alleles are localized within a 1.2-kilobase genomic fragment, of which only transcription unit II is a part. The cDNA of this unit recognizes 1.65- and 1.8-kilobase messages in wild-type Drosophila adult head and body tissues that are absent or extremely reduced in pn mutants. Taken together, the results suggest that transcription unit II defines a part of the pn locus and its cDNA encodes a putative structural gene of pn.

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