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.

REFERENCES

Anderson
K. V.
,
Bokla
L.
,
Nusslein-Volhard
C.
(
1985
).
Establishment of dorsal-ventral polarity in the Drosophila embryo: the induction of polarity by the Toll gene product.
Cell
42
,
791
798
Bonfini
L.
,
Karlovich
C. A.
,
Dasgupta
C.
,
Banerjee
U.
(
1992
).
The Son of sevenless gene product: a putative activator of Ras.
Science
255
,
603
606
Buckles
G. R.
,
Smith
Z. D. J.
,
Katz
F. N.
(
1992
).
mip causes hyperinnervation of a retinotopic map in Drosophila by excessive recruitment of R7 photoreceptor cells.
Neuron
8
,
1015
1029
Busson
D.
,
Gans
M.
,
Komitopoulou
K.
,
Masson
M.
(
1983
).
Genetic analysis of three dominant female sterile mutations located on the X chromosome of Drosophila melanogaster.
Genetics
105
,
309
325
Chou
T.-B.
,
Perrimon
N.
(
1992
).
Use of a yeast site-specific recombinase to produce female germline chimeras in Drosophila.
Genetics
131
,
643
653
Chou
T.-B.
,
Mims
I.
,
Belanich
M.
,
Zachar
Z.
,
Bingham
P. M.
(
1987
).
Procedural improvements in injections for P-mediated germline gene transfer.
Dros. Inf. Serv
66
,
156
157
Clifford
R. T.
,
Schupbach
T.
(
1989
).
Coordinately and differentially mutable activities of torpedo, the Drosophila melanogaster homologue of the vertebrate EGF receptor gene.
Genetics
123
,
771
787
Comer
A. R.
,
Searles
L. L.
,
Kalfayan
L. J.
(
1992
).
Identfication of a genomic DNA fragment containing the Drosophila melanogasterovarian tumor gene (otu) and localization of regions governing its expression.
Gene
118
,
171
179
Erdelyi
M.
,
Szabad
J.
(
1989
).
Isolation and characterization of dominant female sterile mutations of Drosophila melanogaster. I. Mutations on the third chromosome.
Genetics
122
,
111
127
Frank
L. H.
,
Cheung
H.-K.
,
Cohen
R. S.
(
1992
).
Identification and characterization of Drosophila female germ line transcriptional control elements.
Development
114
,
481
491
Freeman
M.
,
Kimmel
B. E.
,
Rubin
G. M.
(
1992
).
Identifying targets of the rough homeobox gene of Drosophila: evidence that rhomboid functions in eye development.
Development
116
,
335
346
Gans
M.
,
Audit
C.
,
Masson
M.
(
1975
).
Isolation and characterization of sex-linked female sterile mutations in Drosophila melanogaster.
Genetics
81
,
683
704
Garfinkel
M. D.
,
Lohe
A. R.
,
Mahowald
A. P.
(
1992
).
Molecular genetics of the Drosophila melanogaster ovo locus, a gene required for sex determination of germline cells.
Genetics
130
,
791
803
Gaul
U.
,
Mardon
G.
,
Rubin
G.
(
1992
).
A putative Ras GTPaseacitivating protein acts as a negative regulator of signaling by the sevenless receptor tyrosine kinase.
Cell
68
,
1007
1019
Golic
K.
,
Lindquist
S.
(
1989
).
The FLP recombinase of yeast catalyzes site-specific recombination in the Drosophila genome.
Cell
59
,
499
509
Golic
K. G.
(
1991
).
Site-specific recombination between homologous chromosomes in Drosophila.
Science
252
,
958
961
Govind
S.
,
Steward
R.
(
1991
).
Dorsoventral pattern formation in Drosophila: Signal transduction and nuclear targeting.
Trend. Gen
7
,
119
125
Haenlin
M.
,
Steller
H.
,
Pirrotta
V.
,
Mohier
E.
(
1985
).
A 43 kilobase cosmid P transposon rescues the fs(1)K10 morphogenetic locus and three adjacent Drosophila developmental mutants.
Cell
40
,
827
837
Hamilton
B. A.
,
Palazzolo
M. J.
,
Meyerowitz
E. M.
(
1991
).
Rapid isolation of long cDNA clones from existing libraries.
Nucleic Acids Res
19
,
1951
1952
Hawley
R. S.
,
Steuber
R. A.
,
Marcus
C. H.
,
Sohn
R.
,
Baronas
D. M.
,
Gameron
M. L.
,
Zitron
A. E.
,
Chase
J. W.
(
1988
).
Molecular analysis of an unstable P element insertion at the singed locus of Drosophila melanogaster evidence for intracistonic transposition of a P element.
Genetics
119
,
85
94
Ish-Horowicz
D.
,
Burke
J. F.
(
1981
).
Rapid and efficient cosmid cloning.
Nucleic Acid Res
9
,
2989
–.
Karess
R. E.
,
Rubin
G. M.
(
1984
).
Analysis of P tranposable element functions in Drosophila.
Cell
38
,
135
146
Kelley
R. L.
(
1993
).
Initial organization of the Drosophila dorsoventral axis depends on an RNA-binding protein encoded by the squid gene.
Genes Dev
7
,
948
960
Kemphues
K. J.
,
Raff
E. C.
,
Raff
R. A.
,
Kaufman
T. C.
(
1980
).
Mutation in a testis-specific β-tublin in Drosophila: analysis of its effects on meiosis and map location of the gene.
Cell
21
,
445
451
Klingler
M.
,
Erdelyi
M.
,
Szabad
J.
,
Nusslein-volhard
C.
(
1988
).
The function of torso in determining the termini anlagen of the Drosophila embryo.
Nature
335
,
275
277
Komitopoulou
K.
,
Gans
M.
,
Margaritis
L. M.
,
Kafatos
F.C.
,
Masson
M.
(
1983
).
Isolation and characterization of sex-linked female-sterile mutations in Drosophila melanogaster with special attention to eggshell mutants.
Genetics
105
,
897
920
Laski
F. A.
,
Rio
D. C.
,
Rubin
G. M.
(
1986
).
Tissue specificity of Drosophila P element transposition is regulated at the level of mRNA splicing.
Cell
44
,
7
19
Laski
F. A.
,
Rubin
G. M.
(
1989
).
Analysis of the cis-acting requirements for germ-line-specific splicing of the P-element ORF2-ORF3 intron.
Genes Dev
3
,
720
728
Lu
X.
,
Chou
T.-B.
,
Willians
N. G.
,
Roberts
T.
,
Perrimon
N.
(
1993
).
Control of cell fate determination by p21ras /Ras1, an essential component of torso signaling in Drosophila.
Genes Dev
7
,
621
632
Manseau
L. J.
,
Schupbach
T.
(
1989
).
Cappuccino and spire: two unique maternal-effect loci required for both the anteroposterior and dorsal-ventral patterns of the Drosophila embryo.
Genes Dev
3
,
1437
1452
MacDonald
P. M.
,
Struhl
G.
(
1986
).
A molecular gradient in early Drosophila embryos and its role in specifying the body pattern.
Nature
324
,
537
545
Mayer
U.
,
Nusslein-Volhard
C.
(
1988
).
A group of genes required for pattern formation in the ventral ectoderm of the Drosophila embryo.
Genes Dev
2
,
1496
1511
Mevel-Ninio
M.
,
Terracol
R.
,
Kafatos
F.C.
(
1991
).
The ovo gene of Drosophila encodes a zinc finger protein required for female germ line development.
EMBOJ
10
,
2259
2266
Mohler
J. D.
(
1977
).
Developmental genetics of the Drosophila egg. I. Identification of 50 sex-linked cistrons with maternal effects on embryonic development.
Genetics
85
,
259
272
Oliver
B.
,
Perrimon
N.
,
Mahowald
A.P.
(
1987
).
The ovo locus is required for sex-specific germ line maintenance in Drosophila.
Genes Dev
1
,
913
923
Oliver
B.
,
Perrimon
N.
,
Mahowald
A.P.
(
1988
).
Genetic evidenceT.-B. Chou, E. Noll and N. Perrimon1369Autosomal dominant female-sterile mutationsthat the sans fille locus is involved in Drosophila sex determination.
Genetics
120
,
159
171
Olivier
J. P.
,
Rabbe
T.
,
Henkemeyer
M.
,
Dickson
B.
,
Mbamalu
G.
,
Margolis
B.
,
Schlessinger
J.
,
Hafen
E.
,
Pawson
T.
(
1993
).
A Drosophila SH2-SH3 adaptor protein implicated in coupling the Sevenless tyrosine kinase to an activatior of Ras Guanine nucleotide exchange, Sos.
Cell
73
,
179
191
Palmiter
R. D.
,
Behringer
R. R.
,
Quaife
C. J.
,
Maxwell
F.
,
Maxwell
I. H.
,
Brinster
R. L.
(
1987
).
Cell lineage ablaation in transgenic mice by cell-specific expression of a toxin gene.
Cell
50
,
435
443
Perrimon
N.
,
Gans
M.
(
1983
).
Clonal analysis of the tissue specificity of recessive female sterile mutations of Drosophila melanogaster using a dominant female sterile mutation Fs(1)K1237.
Dev. Biol
100
,
365
373
Perrimon
N.
(
1984
).
Clonal analysis of dominant female sterile, germline-dependent mutations in Drosophila melanogaster.
Genetics
108
,
927
939
Perrimon
N.
,
Engstrom
L.
,
Mahowald
A.P.
(
1984
).
The effects of zygotic lethal mutations on female germ-line functions in Drosophila.
Dev. Biol
105
,
404
414
Perrimon
N.
,
Mohler
J. D.
,
Engstrom
L.
,
Mahowald
A.P.
(
1986
).
X-linked female sterile loci in Drosophila melanogaster.
Genetics
113
,
695
712
Perrimon
N.
,
Engstrom
L.
,
Mahowald
A. P.
(
1989
).
Zygotic lethals with specific maternal effect phenotypes in Drosophila melanogaster. I. Loci on the X-chromosome.
Genetics
121
,
333
352
Perrimon
N.
(
1993
).
The Torso receptor tyrosine kinase signaling pathway: An endless story.
Cell
, in press.
Price
J. V.
,
Clifford
R. J.
,
Schupbach
R.
(
1989
)
The maternal ventralizing locus torpedo is allelic to faint little ball, an embryonic lethal, and encodes the Drosophila EGF receptor homolog.
Cell
56
,
1085
1092
Robertson
H. M.
,
Preston
C. R.
,
Phillis
R. W.
,
Johnson-Schlitz
D.
,
Benz
W. K.
,
Engels
W. R.
(
1988
).
A stable source of P-element transposase in Drosophila melanogaster.
Genetics
118
,
461
470
Rogge
R. D.
,
Karlovich
C. A.
,
Banerjee
U.
(
1991
).
Genetics dissection of a neurodevelopmental pathway: Son of sevenless functions downstream of the sevenless and EGF receptor tyrosine kinase.
Cell
64
,
39
48
Ruohola-Baker
H.
,
Grell
E.
,
Chou
T.-B.
,
Baker
D.
,
Jan
L. Y.
,
Jan
Y. N.
(
1993
).
Spatially localized Rhomboid is required for establishment of the dorsal-ventral axis in Drosophila oogenesis.
Cell
73
,
953
965
Salz
H. K.
,
Cline
T. W.
,
Schedl
P.
(
1987
).
Functional changes associated with structural alterations induced by mobilization of a P element inserted in the sex-lethal gene of Drosophila.
Genetics
117
,
221
231
Schupbach
T.
,
Wieschaus
E.
(
1986
).
Germline autonomy of maternal-effect mutations altering the embryonic body pattern of Drosophila.
Dev. Biol
113
,
443
448
Schupbach
T.
,
Wieschaus
E.
(
1989
).
Female sterile mutations on the second chromosome of Drosophila melanogaster. I. Maternal effect mutations.
Genetics
121
,
101
117
Schupbach
T.
(
1987
).
Germline and soma cooperate during oogenesis to eatablish the dorso-ventral pattern of egg shell and embryo in Drosophila melanogaster.
Cell
49
,
699
707
Schejter
E. D.
,
Shilo
B.-Z.
(
1989
).
The Drosophila EGF recptor homolog (DER) gene is allelic to faint little ball, a locus essential for embryonic development.
Cell
56
,
1093
1104
Simon
A. M.
,
Bowtell
D. D. L.
,
Dodson
G. S.
,
Laverty
T. R.
,
Rubin
G. M.
(
1991
).
Ras1 and a putative guanine nucleotide exchange factor perform crucial steps in signaling by the sevenless protein tyrosine kinase.
Cell
67
,
701
716
Simon
A. M.
,
Dodson
G.S.
,
Rubin
G. M.
(
1993
).
An SH3-SH2-SH3 protein is required for p21Ras1 activation and binds to Sevenless and Sos proteins in vitro.
Cell
73
,
169
177
Sosnowski
B. A.
,
Belote
J. M.
,
McKeown
M.
(
1989
).
Sex-specific, alternative splicing of RNA from the transformer gene results from sequence-dependent splice site blockage.
Cell
58
,
449
459
Sturtevant
M. A.
,
Roark
M.
,
Bier
E.
(
1993
).
The Drosophila rhomboid gene mediates the localized formation of wing veins and interacts genetically with components of the EGF-R signaling pathway.
Genes Dev
7
,
961
973
Szabad
J.
,
Erdelyi
M.
,
Szidonya
J.
(
1987
).
Characterization of Fs(2)1, a germ line dependent dominant female sterile mutation of Drosophila.
Acta Biol. Hung
38
,
257
266
Szabad
J.
,
Erdelyi
M. G.
,
Hoffmann
M. G.
,
Szidonya
J.
,
Wright
T. R. F.
(
1989
).
Isolation and characterization of dominant female sterile mutations of Drosophila melanogaster. II. Mutations on the second chromosome.
Genetics
122
,
823
835
Thummel
C. S.
,
Pirrotta
V.
(
1992
).
New pCaSpeR P element vectors.
Dros. Inf. Service
71
,
150
–.
Tower
J.
,
Karpen
G. H.
,
Craig
N.
,
Spradling
A. C.
(
1993
).
Preferential transposition of Drosophila P elements to nearby chromosomal sites.
Genetics
133
,
347
359
van der Meer
J.
(
1977
).
Optical clean and permanent whole mount preparations for phase contrast microscopy of cuticular structures of insect larvae.
Dros. Inf. Serv
52
,
160
–.
Wieschaus
E.
,
Szabad
J.
(
1979
).
The developmental and function of the female germ line in Drosophila melanogaster: A cell lineage study.
Dev. Biol
68
,
29
46
Wieschaus
E.
,
Audit
C.
,
Masson
M.
(
1981
).
A clonal analysis of the roles of somatic cells and germ line during oogenesis in Drosophila.
Dev. Biol
88
,
92
103
Wieschaus
E.
,
Nusslein-Volhard
C.
,
Kluding
H.
(
1984
).
Kruppel, a gene whose activity is required early in the zygotic genome for normal embryonic segmentation.
Dev. Biol
104
,
172
186
Yarger
R. G.
,
King
R. C.
(
1971
).
The phenogenetics of a temperature sensitive, autosomal dominant, female sterile gene in Drosophila melanogaster.
Dev. Biol
24
,
166
177
Zhang
P.
,
Spradling
A. C.
(
1993
).
Efficient and dispersed local P element transposition from Drosophila females.
Genetics
133
,
361
373
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