Restriction of P-element insertions at the Notch locus of Drosophila melanogaster

P elements move about the Drosophila melanogaster genome in a nonrandom fashion, preferring some chromosomal targets for insertion over others (J. C. J. Eeken, F. H. Sobels, V. Hyland, and A. P. Schalet, Mutat. Res. 150:261-275, 1985; W. R. Engels, Annu. Rev. Genet. 17:315-344, 1983; M. D. Golubovsky, Y. N. Ivanov, and M. M. Green, Proc. Natl. Acad. Sci. USA 74:2973-2975, 1977; M. J. Simmons and J. K. Lim, Proc. Natl. Acad. Sci. USA 77:6042-6046, 1980). Some of this specificity may be due to recognition of a particular DNA sequence in the target DNA; derivatives of an 8-base-pair consensus sequence are occupied by these transposable elements at many different chromosomal locations (K. O'Hare and G. M. Rubin, Cell 34:25-36, 1983). An additional level of specificity of P-element insertions is described in this paper. Of 14 mutations induced in the complex locus Notch by hybrid dysgenesis, 13 involved P-element insertions at or near the transcription start site of the gene. This clustering was not seen in other transposable element-induced mutations of Notch. DNA sequences homologous to the previously described consensus target for P-element insertion are not preferentially located in this region of the locus. The choice of a chromosomal site for integration appears to be based on more subtle variations in chromosome structure that are probably associated with activation or expression of the target gene.

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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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The molecular organization of the Star/asteroid region, a region necessary for proper eye development in Drosophila melanogaster

Genome ◽

10.1139/g93-049 ◽

1993 ◽

Vol 36 (2) ◽

pp. 356-366 ◽

Cited By ~ 4

Author(s):

Theodore S. Higson ◽

June E. Tessiatore ◽

Sean A. Bennett ◽

Raymond C. Derk ◽

Michael A. Kotarski

Keyword(s):

Drosophila Melanogaster ◽

Molecular Data ◽

P Element ◽

Molecular Organization ◽

Induced Mutations ◽

Dna Breaks ◽

X Ray ◽

P Elements ◽

Large Deletions ◽

Endonuclease Mapping

The Star/asteroid (S/ast) region of Drosophila melanogaster has been cloned by P element transposon tagging using the snw chromosome as a source of defective P elements. In each mutation examined, the element integrated into the region was a 0.5-kb element from a region proximal to sn and not one of the head-to-head elements from the sn locus. Previously described spontaneous and X-ray induced mutations of S and ast were located on the molecular map by Southern analysis and restriction endonuclease mapping of genomic clones. S mutations are either large deletions of the cloned region or DNA breaks located near the P element insertions that cause ast mutations. Both S and ast mutations reduce the steady-state amounts of a 3.4-kb RNA. The molecular data, together with the phenotypic interactions observed for S and ast alleles, are consistent with the interpretation that S and ast mutations are lesions within the same gene or within genes that are functionally related.Key words: Drosophila, Star, asteroid, P elements.

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

Molecular and Cellular Biology ◽

10.1128/mcb.6.5.1520-1528.1986 ◽

1986 ◽

Vol 6 (5) ◽

pp. 1520-1528

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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Transposable elements and fitness in Drosophila melanogaster

Genome ◽

10.1139/g89-046 ◽

1989 ◽

Vol 31 (1) ◽

pp. 284-295 ◽

Cited By ~ 35

Author(s):

Trudy F. C. Mackay

Keyword(s):

Drosophila Melanogaster ◽

Transposable Elements ◽

Insertional Mutagenesis ◽

Natural Populations ◽

P Element ◽

Induced Mutations ◽

P Elements ◽

Copy Numbers ◽

Genomic Locations

Transposable elements constitute a significant fraction of the Drosophila melanogaster genome. The five families of moderately repeated transposable elements identified to date occupy dispersed and variable genomic locations, but have relatively constant copy numbers per individual. What effect to these elements have on the fitness of the individuals harboring them? Experimental evidence relating to this question is reviewed. The relevant data fall into two broad categories. The first involves the determination of the distribution of transposable elements in natural populations, by restriction mapping or in situ hybridization, and the comparison of the observed distribution with different theoretical expectations. The second approach is to study directly the effects of new transposable element-induced mutations on fitness. The P family of transposable elements is a particularly efficient mutagen, and the results of experiments in which initially P-free chromosomes are contaminated with P elements are discussed with regard to P-induced fitness mutations.Key words: transposable elements, Drosophila melanogaster, insertional mutagenesis, fitness, P element mutagenesis, hybrid dysgenesis.

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P-Element Insertion Alleles of Essential Genes on the Third Chromosome of Drosophila melanogaster Correlation of Physical and Cytogenetic Maps in Chromosomal Region 86E-87F

Genetics ◽

10.1093/genetics/147.4.1697 ◽

1997 ◽

Vol 147 (4) ◽

pp. 1697-1722 ◽

Cited By ~ 1

Author(s):

Peter Deák ◽

Mahmoud M Omar ◽

Robert D C Saunders ◽

Margit Pál ◽

Orbán Komonyi ◽

...

Keyword(s):

Drosophila Melanogaster ◽

Genetic Maps ◽

P Element ◽

Drosophila Genome ◽

Induced Mutations ◽

Lethal Mutant ◽

P Elements ◽

Element Insertion ◽

The Third ◽

Complementation Groups

Abstract We have established a collection of 2460 lethal or semi-lethal mutant lines using a procedure thought to insert single P elements into vital genes on the third chromosome of Drosophila melanogaster. More than 1200 randomly selected lines were examined by in situ hybridization and 90% found to contain single insertions at sites that mark 89% of all lettered subdivisions of the Bridges' map. A set of chromosomal deficiencies that collectively uncover ~25% of the euchromatin of chromosome 3 reveal lethal mutations in 468 lines corresponding to 145 complementation groups. We undertook a detailed analysis of the cytogenetic interval 86E-87F and identified 87 P-element-induced mutations falling into 38 complementation groups, 16 of which correspond to previously known genes. Twenty-one of these 38 complementation groups have at least one allele that has a P-element insertion at a position consistent with the cytogenetics of the locus. We have rescued P elements and flanking chromosomal sequences from the 86E-87F region in 35 lines with either lethal or genetically silent P insertions, and used these as probes to identify cosmids and P1 clones from the Drosophila genome projects. This has tied together the physical and genetic maps and has linked 44 previously identified cosmid contigs into seven “super-contigs” that span the interval. STS data for sequences flanking one side of the P-element insertions in 49 lines has identified insertions in the αγ element at 87C, two known transposable elements, and the open reading frames of seven putative single copy genes. These correspond to five known genes in this interval, and two genes identified by the homology of their predicted products to known proteins from other organisms.

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

Genetics ◽

10.1093/genetics/161.1.195 ◽

2002 ◽

Vol 161 (1) ◽

pp. 195-204 ◽

Cited By ~ 1

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.

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The Regulatory Properties of Autonomous Subtelomeric P Elements Are Sensitive to a Suppressor of variegation in Drosophila melanogaster

Genetics ◽

10.1093/genetics/143.4.1663 ◽

1996 ◽

Vol 143 (4) ◽

pp. 1663-1674 ◽

Cited By ~ 3

Author(s):

Stéphane Ronsseray ◽

Monique Lehmann ◽

Danielle Nouaud ◽

Dominique Anxolabéhère

Keyword(s):

Drosophila Melanogaster ◽

Genetic Recombination ◽

Natural Populations ◽

P Element ◽

Single Element ◽

Heterochromatin Protein 1 ◽

X Chromosomes ◽

P Elements ◽

Associated Sequences ◽

Regulatory Properties

Abstract Genetic recombination was used in Drosophila melanogaster to isolate P elements, inserted at the telomeres of X chromosomes (cytological site 1A) from natural populations, in a genetic background devoid of other P elements. We show that complete maternally inherited P repression in the germline (P cytotype) can be elicited by only two autonomous P elements at 1A and that a single element at this site has partial regulatory properties. The analysis of the surrounding chromosomal regions of the P elements at 1A shows that in all cases these elements are flanked by Telomeric Associated Sequences, tandemly repetitive noncoding sequences that have properties of heterochromatin. In addition, we show that the regulatory properties of P elements at 1A can be inhibited by some of the mutant alleles of the Su(var)205 gene and by a deficiency of this gene. However, the regulatory properties of reference P strains (Harwich and Texas 007) are not impaired by Su(var)205 mutations. Su(var)205 encodes Heterochromatin Protein 1 (HP1). These results suggest that the HP1 dosage effect on the P element properties is sitedependent and could involve the structure of the chromatin.

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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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A novel repressor of P element transposition in Drosophila melanogaster

Genetics Research ◽

10.1017/s0016672397003066 ◽

1998 ◽

Vol 71 (1) ◽

pp. 21-30 ◽

Cited By ~ 4

Author(s):

RICHARD M. BADGE ◽

JOHN F. Y. BROOKFIELD

Keyword(s):

Drosophila Melanogaster ◽

Inbred Line ◽

Gonadal Dysgenesis ◽

Full Length ◽

P Element ◽

Temperature Dependent ◽

P Elements

We have discovered, in an inbred line (Loua) of Drosophila melanogaster from Zaïre, a third chromosome showing unusual P element repression. Repression of P element transposition by this chromosome, named Loua3, is dominant zygotic and has three unusual properties. Firstly, its repression of the gonadal dysgenesis caused by a strong P haplotype is strongly temperature-dependent, being most evident at higher rearing temperatures. Secondly, subdivision of Loua3 by recombination abolishes repression: the effect is apparently a function of the intact chromosome. Finally, Loua3 also diminishes somatic lethality when chromosomes carrying many ‘ammunition’ elements (Birmingham2) are exposed to the constitutive transposase source Δ2-3(99B). The chromosome has 17 P elements, none full-length, located in at least 12 dispersed positions.

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