A New Resource for Characterizing X-Linked Genes in Drosophila melanogaster: Systematic Coverage and Subdivision of the X Chromosome With Nested, Y-Linked Duplications

Interchromosomal duplications are especially important for the study of X-linked genes. Males inheriting a mutation in a vital X-linked gene cannot survive unless there is a wild-type copy of the gene duplicated elsewhere in the genome. Rescuing the lethality of an X-linked mutation with a duplication allows the mutation to be used experimentally in complementation tests and other genetic crosses and it maps the mutated gene to a defined chromosomal region. Duplications can also be used to screen for dosage-dependent enhancers and suppressors of mutant phenotypes as a way to identify genes involved in the same biological process. We describe an ongoing project in Drosophila melanogaster to generate comprehensive coverage and extensive breakpoint subdivision of the X chromosome with megabase-scale X segments borne on Y chromosomes. The in vivo method involves the creation of X inversions on attached-XY chromosomes by FLP-FRT site-specific recombination technology followed by irradiation to induce large internal X deletions. The resulting chromosomes consist of the X tip, a medial X segment placed near the tip by an inversion, and a full Y. A nested set of medial duplicated segments is derived from each inversion precursor. We have constructed a set of inversions on attached-XY chromosomes that enable us to isolate nested duplicated segments from all X regions. To date, our screens have provided a minimum of 78% X coverage with duplication breakpoints spaced a median of nine genes apart. These duplication chromosomes will be valuable resources for rescuing and mapping X-linked mutations and identifying dosage-dependent modifiers of mutant phenotypes.

[1]  E. Popodi,et al.  A Molecularly Defined Duplication Set for the X Chromosome of Drosophila melanogaster , 2010, Genetics.

[2]  Jiangang Gao,et al.  A review of current large‐scale mouse knockout efforts , 2010, Genesis.

[3]  J. Kumar,et al.  The Retinal Determination Gene eyes absent Is Regulated by the EGF Receptor Pathway Throughout Development in Drosophila , 2010, Genetics.

[4]  Norbert Perrimon,et al.  A Drosophila Resource of Transgenic RNAi Lines for Neurogenetics , 2009, Genetics.

[5]  Yves Moreau,et al.  Integrating Computational Biology and Forward Genetics in Drosophila , 2009, PLoS genetics.

[6]  David Osumi-Sutherland,et al.  FlyBase: enhancing Drosophila Gene Ontology annotations , 2008, Nucleic Acids Res..

[7]  P. Szafranski,et al.  Basolateral Junctions Utilize Warts Signaling to Control Epithelial-Mesenchymal Transition and Proliferation Crucial For Migration and Invasion of Drosophila Ovarian Epithelial Cells , 2008, Genetics.

[8]  Michael Ashburner,et al.  The DrosDel Deletion Collection: A Drosophila Genomewide Chromosomal Deficiency Resource , 2007, Genetics.

[9]  B. Dickson,et al.  A genome-wide transgenic RNAi library for conditional gene inactivation in Drosophila , 2007, Nature.

[10]  M. Leptin,et al.  Analysis and reconstitution of the genetic cascade controlling early mesoderm morphogenesis in the Drosophila embryo , 2007, Mechanisms of Development.

[11]  Hugo J. Bellen,et al.  P[acman]: A BAC Transgenic Platform for Targeted Insertion of Large DNA Fragments in D. melanogaster , 2006, Science.

[12]  R. Hawley,et al.  The Mechanism of Secondary Nondisjunction in Drosophila melanogaster Females , 2006, Genetics.

[13]  G. Rubin,et al.  The BDGP Gene Disruption Project , 2004, Genetics.

[14]  M. Ashburner,et al.  The DrosDel Collection , 2004, Genetics.

[15]  D. Curtis,et al.  Systematic generation of high-resolution deletion coverage of the Drosophila melanogaster genome , 2004, Nature Genetics.

[16]  Feng Chen,et al.  A complementary transposon tool kit for Drosophila melanogaster using P and piggyBac , 2004, Nature Genetics.

[17]  Shigeo Hayashi,et al.  GETDB, a database compiling expression patterns and molecular locations of a collection of gal4 enhancer traps , 2002, Genesis.

[18]  K. Golic,et al.  Engineering the Drosophila genome: chromosome rearrangements by design. , 1996, Genetics.

[19]  D. Lindsley,et al.  The Genome of Drosophila Melanogaster , 1992 .

[20]  H. Salz.  The genetic analysis of snf: a Drosophila sex determination gene required for activation of Sex-lethal in both the germline and the soma. , 1992, Genetics.

[21]  W. Engels,et al.  High-frequency P element loss in Drosophila is homolog dependent , 1990, Cell.

[22]  Michael Ashburner,et al.  Drosophila: A laboratory handbook , 1990 .

[23]  J. Kennison.  The Genetic and Cytological Organization of the Y Chromosome of DROSOPHILA MELANOGASTER. , 1981, Genetics.

[24]  S. Zimmering,et al.  MEIOTIC X-Y EXCHANGE AND NONDISJUNCTION INDUCED BY IRRADIATION IN THE DROSOPHILA MALE. , 1964, Genetics.

[25]  E. Sutton.  A Cytogenetic Study of the Yellow-Scute Region of the X Chromosome in Drosophila Melanogaster. , 1943, Genetics.

[26]  J. Patterson,et al.  Further Studies on X Chromosome Balance in Drosophila. , 1937, Genetics.

[27]  E. Spana,et al.  Mapping Drosophila genomic aberration breakpoints with comparative genome hybridization on microarrays. , 2006, Methods in enzymology.

[28]  A.T.C. Carpenter.  Salivary chromosome analysis of aberrations. , 2004, Methods in molecular biology.

[29]  L. Fanti,et al.  Analysis of mitosis in squash preparations of larval brains: orcein, Giemsa, Hoechst 33258, DAPI, quinacrine, and N-banding. , 2004, Methods in molecular biology.