title: A genetic screen based on in vivo RNA imaging reveals centrosome-independent mechanisms for localizing gurken transcripts in Drosophila.
abstract: We have screened chromosome arm 3L for ethyl methanesulfonate-induced mutations that disrupt localization of fluorescently labeled gurken (grk) messenger (m)RNA, whose transport along microtubules establishes both major body axes of the developing Drosophila oocyte. Rapid identification of causative mutations by single-nucleotide polymorphism recombinational mapping and whole-genomic sequencing allowed us to define nine complementation groups affecting grk mRNA localization and other aspects of oogenesis, including alleles of elg1, scaf6, quemao, nudE, Tsc2/gigas, rasp, and Chd5/Wrb, and several null alleles of the armitage Piwi-pathway gene. Analysis of a newly induced kinesin light chain allele shows that kinesin motor activity is required for both efficient grk mRNA localization and oocyte centrosome integrity. We also show that initiation of the dorsoanterior localization of grk mRNA precedes centrosome localization, suggesting that microtubule self-organization contributes to breaking axial symmetry to generate a unique dorsoventral axis.
authors: Developmental Genetics Laboratory and.
authors: Rippei
authors: R
authors: Hayashi
authors: S Mark
authors: SM
authors: Wainwright
authors: Sophie J
authors: SJ
authors: Liddell
authors: Sheena M
authors: SM
authors: Pinchin
authors: Stuart
authors: S
authors: Horswell
authors: David
authors: D
authors: Ish-Horowicz
mesh_terms: D000818:Animals; D018385:Centrosome / Q000378:metabolism*; D002874:Chromosome Mapping; D004330:Drosophila / Q000254:growth & development / Q000378:metabolism*; D029721:Drosophila Proteins / Q000037:antagonists & inhibitors / Q000235:genetics* / Q000378:metabolism; D005260:Female; D005820:Genetic Testing; D059014:High-Throughput Nucleotide Sequencing; D016547:Kinesins / Q000235:genetics / Q000378:metabolism; D008297:Male; D009865:Oocytes / Q000378:metabolism; D009866:Oogenesis; D020641:Polymorphism, Single Nucleotide; D012313:RNA / Q000737:chemistry / Q000378:metabolism*; D012333:RNA, Messenger / Q000032:analysis / Q000378:metabolism; D034741:RNA, Small Interfering / Q000378:metabolism; D017422:Sequence Analysis, DNA; D016211:Transforming Growth Factor alpha / Q000235:genetics* / Q000378:metabolism
chemical_list: D029721:Drosophila Proteins; D012333:RNA, Messenger; D034741:RNA, Small Interfering; D016211:Transforming Growth Factor alpha; C083730:grk protein, Drosophila; D012313:RNA; D016547:Kinesins
keywords: RNA localization; microtubules; mutant mapping; oogenesis; whole-genome sequencing