Sites of P element insertion and structures of P element deletions in the 5' region of Drosophila melanogaster RpII215.

Several P element insertion and deletion mutations near the 5' end of Drosophila melanogaster RpII215 have been examined by nucleotide sequencing. Two different sites of P element insertion, approximately 90 nucleotides apart, have been detected in this region of the gene. Therefore, including an additional site of P element insertion within the coding region, there are at least three distinct sites of P element insertion at RpII215. Both 5' sites are within a noncoding portion of transcribed sequences. The sequences of four revertants of one P element insertion mutation (D50) indicate that the P element is either precisely deleted or internally deleted to restore RpII215 activity. Partial internal deletions of the P element result in different RpII215 activity levels, which appear to depend on the specific sequences that remain after excision.

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Hsp28stl: A P-Element Insertion Mutation That Alters the Expression of a Heat Shock Gene in Drosophila melanogaster

Genetics ◽

10.1093/genetics/115.2.333 ◽

1987 ◽

Vol 115 (2) ◽

pp. 333-340

Author(s):

Joel C Eissenberg ◽

Sarah C R Elgin

Keyword(s):

Drosophila Melanogaster ◽

Heat Shock ◽

P Element ◽

Developmental Expression ◽

Heat Shock Gene ◽

Insertion Mutation ◽

Target Sequence ◽

Element Insertion ◽

Large Heat ◽

Shock Gene

ABSTRACT We have identified and cloned a mutant allele of the small heat shock gene Hsp28 of Drosophila melanogaster. This allele, which we have called Hsp28stl, produces small amounts of a single aberrantly large, heat-inducible transcript in heat-shocked flies, while a normal-sized Hsp28 transcript is present only in fertile females. No Hsp28 transcript at all is detected in mutant prepupae, a stage when wild-type flies show high levels of Hsp28 RNA. We have cloned the Hsp28stl allele, and have found that a 1.3-kb defective P-element is present 5' to Hsp28 in the mutant line. The site of P-element insertion lies between the Hsp28 "TATA box" sequence and the Hsp28 RNA cap site; in contrast to previously described P-element insertions, the element at Hsp28stl is flanked by a two base pair duplication of the insertional target sequence. The results suggest that this insert may separate elements regulating heat-inducible and developmental expression of Hsp28, leading to the different patterns of transcription observed.

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The effect of X-irradiation and formaldehyde treatment of spermatogonia on the reversion of an unstable, P-element insertion mutation in Drosophila melanogaster

Mutation Research Letters ◽

10.1016/0165-7992(86)90126-0 ◽

1986 ◽

Vol 175 (2) ◽

pp. 61-65 ◽

Cited By ~ 9

Author(s):

J.C.J. Eeken ◽

F.H. Sobels

Keyword(s):

Drosophila Melanogaster ◽

P Element ◽

Insertion Mutation ◽

Element Insertion ◽

Formaldehyde Treatment ◽

X Irradiation

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hobo enhancer trapping mutagenesis in Drosophila reveals an insertion specificity different from P elements.

Genetics ◽

10.1093/genetics/135.4.1063 ◽

1993 ◽

Vol 135 (4) ◽

pp. 1063-1076 ◽

Cited By ~ 2

Author(s):

D Smith ◽

J Wohlgemuth ◽

B R Calvi ◽

I Franklin ◽

W M Gelbart

Keyword(s):

Drosophila Melanogaster ◽

Transposable Element ◽

Enhancer Trap ◽

P Element ◽

Present Evidence ◽

P Elements ◽

Element Insertion ◽

Enhancer Trapping ◽

Indispensable Tool

Abstract P element enhancer trapping has become an indispensable tool in the analysis of the Drosophila melanogaster genome. However, there is great variation in the mutability of loci by these elements such that some loci are relatively refractory to insertion. We have developed the hobo transposable element for use in enhancer trapping and we describe the results of a hobo enhancer trap screen. In addition, we present evidence that a hobo enhancer trap element has a pattern of insertion into the genome that is different from the distribution of P elements in the available database. Hence, hobo insertion may facilitate access to genes resistant to P element insertion.

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Molecular organization and expression of the genetic locus glued in Drosophila melanogaster

Molecular and Cellular Biology ◽

10.1128/mcb.6.3.833-841.1986 ◽

1986 ◽

Vol 6 (3) ◽

pp. 833-841

Author(s):

A Swaroop ◽

J W Sun ◽

M L Paco-Larson ◽

A Garen

Keyword(s):

Drosophila Melanogaster ◽

Cell Function ◽

Genetic Locus ◽

Insertion Site ◽

Lethal Effect ◽

P Element ◽

Molecular Organization ◽

General Distribution ◽

Element Insertion ◽

Mutant Phenotypes

The Glued locus of Drosophila melanogaster is genetically defined as the functional unit which is affected by the dominant Glued mutation Gl. Genomic DNA was cloned from the region of the Glued locus, at 70C2 on chromosome 3, by using a P element insertion in the region as a molecular marker. Three genes encoding polyadenylated transcripts were detected within a 30-kilobase span of the cloned DNA. The gene nearest the P element insertion site was identified as a Glued gene on the basis of alterations in its DNA and encoded transcript associated with the Gl mutation and with reversions of Gl which eliminate the dominant effect by inactivation of the mutant allele. Expression of the wild-type Gl+ gene is temporally regulated during development; the amount of the encoded transcript is highest in the embryonic stage, decreasing in the first and second larval instars, and then increasing in the third instar and pupal stages. There is a maternal contribution of the Gl+ transcript to the embryo, which probably accounts for the maternal lethal effect of Glued mutations on early development. In situ hybridizations of Gl+ DNA to RNA in tissue sections showed that the Gl+ transcript is present in virtually all tissues of the embryo, late larva, and pupa. The general distribution of this transcript is consistent with genetic evidence indicating that the Glued locus controls a generally essential cell function (P. J. Harte and D. R. Kankel, Genetics 101:477-501, 1982). Different Glued mutations produce distinct phenotypic effects, including adults with severe visual defects, larvae lacking imaginal discs, and early lethality. These diverse mutant phenotypes are discussed in terms of quantitative changes in the Glued function. Closely adjacent to Gl+ is another gene which is transcribed in a divergent direction and expressed coordinately with Gl+ throughout Drosophila development. It remains to be determined whether this gene is also involved with the Glued function.

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A particular P-element insertion is correlated to the P-induced hybrid dysgenesis repression in Drosophila melanogaster

Genetics Research ◽

10.1017/s0016672300030627 ◽

1992 ◽

Vol 60 (1) ◽

pp. 15-24 ◽

Cited By ~ 5

Author(s):

Dominique Higuet ◽

Dominique Anxolabéhére ◽

Danielle Nouaud

Keyword(s):

Drosophila Melanogaster ◽

Promoter Activity ◽

Hybrid Dysgenesis ◽

P Element ◽

P Elements ◽

Element Insertion ◽

Element Number

SummaryTransposable P elements in Drosophila melanogaster cause hybrid dysgenesis if their mobility is not repressed. The ability to regulate the dysgenic activity of the P elements depends on several mechanisms, one of which hypothesized that a particular deleted P element (the KP element) results in a non-susceptibility which is biparentally transmitted. In this study totally nonsusceptible lines, and susceptible lines containing exclusively KP elements (IINS2 line and IIS2 line) were isolated from a M' strain. We show that non-susceptibility is correlated with a particular insertion of one KP element located at the cytological site 47D1. The repression ability of the GD sterility is determined by a recessive chromosomal factor, and cannot be due to the KP-element number. Here the repression of the P mobility is associated with reduction of the P transcripts and the inhibition of P promoter activity.

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The Gene Search System: A Method for Efficient Detection and Rapid Molecular Identification of Genes in Drosophila melanogaster

Genetics ◽

10.1093/genetics/151.2.725 ◽

1999 ◽

Vol 151 (2) ◽

pp. 725-737 ◽

Cited By ~ 6

Author(s):

Gakuta Toba ◽

Takashi Ohsako ◽

Naomasa Miyata ◽

Tsuyoshi Ohtsuka ◽

Ki-Hyeon Seong ◽

...

Keyword(s):

Drosophila Melanogaster ◽

Core Promoter ◽

Sequence Similarity ◽

Direct Sequencing ◽

P Element ◽

Drosophila Genome ◽

Coding Region ◽

New Genes ◽

Gene Search ◽

Efficient Detection

Abstract We have constructed a P-element-based gene search vector for efficient detection of genes in Drosophila melanogaster. The vector contains two copies of the upstream activating sequence (UAS) enhancer adjacent to a core promoter, one copy near the terminal inverted repeats at each end of the vector, and oriented to direct transcription outward. Genes were detected on the basis of phenotypic changes caused by GAL4-dependent forced expression of vector-flanking DNA, and the transcripts were identified with reverse transcriptase PCR (RT-PCR) using the vector-specific primer and followed by direct sequencing. The system had a greater sensitivity than those already in use for gain-of-function screening: 64% of the vector insertion lines (394/613) showed phenotypes with forced expression of vector-flanking DNA, such as lethality or defects in adult structure. Molecular analysis of 170 randomly selected insertions with forced expression phenotypes revealed that 21% matched the sequences of cloned genes, and 18% matched reported expressed sequence tags (ESTs). Of the insertions in cloned genes, 83% were upstream of the protein-coding region. We discovered two new genes that showed sequence similarity to human genes, Ras-related protein 2 and microsomal glutathione S-transferase. The system can be useful as a tool for the functional mapping of the Drosophila genome.

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Molecular cloning of suppressor of sable, a Drosophila melanogaster transposon-mediated suppressor.

Molecular and Cellular Biology ◽

10.1128/mcb.6.5.1520 ◽

1986 ◽

Vol 6 (5) ◽

pp. 1520-1528 ◽

Cited By ~ 14

Author(s):

D Y Chang ◽

B Wisely ◽

S M Huang ◽

R A Voelker

Keyword(s):

Drosophila Melanogaster ◽

Dna Sequences ◽

Insertion Site ◽

Hybrid Dysgenesis ◽

P Element ◽

Dna Transformation ◽

Wild Type ◽

Induced Mutations ◽

X Ray ◽

Element Insertion

A hybrid dysgenesis-induced allele [su(s)w20] associated with a P-element insertion was used to clone sequences from the su(s) region of Drosophila melanogaster by means of the transposon-tagging technique. Cloned sequences were used to probe restriction enzyme-digested DNAs from 22 other su(s) mutations. None of three X-ray-induced or six ethyl methanesulfonate-induced su(s) mutations possessed detectable variation. Seven spontaneous, four hybrid dysgenesis-induced, and two DNA transformation-induced mutations were associated with insertions within 2.0 kilobases (kb) of the su(s)w20 P-element insertion site. When the region of DNA that included the mutational insertions was used to probe poly(A)+ RNAs, a 5-kb message was detected in wild-type RNA that was present in greatly reduced amounts in two su(s) mutations. By using strand-specific probes, the direction of transcription of the 5-kb message was determined. The mutational insertions lie in DNA sequences near the 5' end of the 5-kb message. Three of the seven spontaneous su(s) mutations are associated with gypsy insertions, but they are not suppressible by su(Hw).

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A selective screen to recover chromosomal deletions and duplications in Drosophila melanogaster.

Genetics ◽

10.1093/genetics/119.2.377 ◽

1988 ◽

Vol 119 (2) ◽

pp. 377-390

Author(s):

D Gubb ◽

S McGill ◽

M Ashburner

Keyword(s):

Drosophila Melanogaster ◽

Chromosomal Region ◽

Molecular Level ◽

Hybrid Dysgenesis ◽

P Element ◽

Genomic Clones ◽

Element Insertion ◽

Chromosomal Deletions ◽

Complementation Groups ◽

Insertion Sites

Abstract A screen is described that will select for breakpoints within a restricted chromosomal region in Drosophila. The aberrations recovered can be used to construct chromosomes carrying synthetic duplications and deletions. Such chromosomes have applications in the mapping of complementation groups at both the genetic and molecular level. In particular, breakpoints recovered after P element hybrid dysgenesis tend to be associated with P element insertion sites. Such aberration breakpoints can be genetically mapped, as synthetic deletions, and then used as transposon-tagged sites for the recovery of genomic clones.

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Structure and expression of hybrid dysgenesis-induced alleles of the ovarian tumor (otu) gene in Drosophila melanogaster.

Genetics ◽

10.1093/genetics/133.2.253 ◽

1993 ◽

Vol 133 (2) ◽

pp. 253-263

Author(s):

G L Sass ◽

J D Mohler ◽

R C Walsh ◽

L J Kalfayan ◽

L L Searles

Keyword(s):

Drosophila Melanogaster ◽

Ovarian Tumor ◽

P Element ◽

Female Sterility ◽

Protein Coding ◽

Transcription Start Sites ◽

Insertion And Deletion ◽

Undifferentiated Cells ◽

Egg Chambers ◽

Mutant Phenotypes

Abstract Mutations at the ovarian tumor (otu) gene of Drosophila melanogaster cause female sterility and generate a range of ovarian phenotypes. Quiescent (QUI) mutants exhibit reduced germ cell proliferation; in oncogenic (ONC) mutants germ cells undergo uncontrolled proliferation generating excessive numbers of undifferentiated cells; the egg chambers of differentiated (DIF) mutants differentiate to variable degrees but fail to complete oogenesis. We have examined mutations caused by insertion and deletion of P elements at the otu gene. The P element insertion sites are upstream of the major otu transcription start sites. In deletion derivatives, the P element, regulatory regions and/or protein coding sequences have been removed. In both insertion and deletion mutants, the level of otu expression correlates directly with the severity of the phenotype: the absence of otu function produces the most severe QUI phenotype while the ONC mutants express lower levels of otu than those which are DIF. The results of this study demonstrate that the diverse mutant phenotypes of otu are the consequence of different levels of otu function.

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