title: A large-scale in vivo RNAi screen to identify genes involved in Notch-mediated follicle cell differentiation and cell cycle switches.
abstract: During Drosophila oogenesis, follicle cells sequentially undergo three distinct cell-cycle programs: the mitotic cycle, endocycle, and gene amplification. Notch signaling plays a central role in regulating follicle-cell differentiation and cell-cycle switches; its activation is essential for the mitotic cycle/endocycle (M/E) switch. Cut, a linker between Notch signaling and cell-cycle regulators, is specifically downregulated by Notch during the endocycle stage. To determine how signaling pathways coordinate during the M/E switch and to identify novel genes involved in follicle cell differentiation, we performed an in vivo RNAi screen through induced knockdown of gene expression and examination of Cut expression in follicle cells. We screened 2205 RNAi lines and found 33 genes regulating Cut expression during the M/E switch. These genes were confirmed with the staining of two other Notch signaling downstream factors, Hindsight and Broad, and validated with multiple independent RNAi lines. We applied gene ontology software to find enriched biological meaning and compared our results with other publications to find conserved genes across tissues. Specifically, we found earlier endocycle entry in anterior follicle cells than those in the posterior, identified that the insulin-PI3K pathway participates in the precise M/E switch, and suggested Nejire as a cofactor of Notch signaling during oogenesis.
authors.affiliation: Department of Biological Science, Florida State University, Tallahassee, FL 32306-4370, USA.
authors.forename: Dongyu
authors.initials: D
authors.lastname: Jia
authors.affiliation: Department of Biological Science, Florida State University, Tallahassee, FL 32306-4370, USA.
authors.forename: Muhammed
authors.initials: M
authors.lastname: Soylemez
authors.affiliation: Department of Biological Science, Florida State University, Tallahassee, FL 32306-4370, USA.
authors.forename: Gabriel
authors.initials: G
authors.lastname: Calvin
authors.affiliation: Department of Biological Science, Florida State University, Tallahassee, FL 32306-4370, USA.
authors.forename: Randy
authors.initials: R
authors.lastname: Bornmann
authors.affiliation: Department of Biological Science, Florida State University, Tallahassee, FL 32306-4370, USA.
authors.forename: Jamal
authors.initials: J
authors.lastname: Bryant
authors.affiliation: Department of Biological Science, Florida State University, Tallahassee, FL 32306-4370, USA.
authors.forename: Cameron
authors.initials: C
authors.lastname: Hanna
authors.affiliation: Department of Biological Science, Florida State University, Tallahassee, FL 32306-4370, USA.
authors.forename: Yi-Chun
authors.initials: YC
authors.lastname: Huang
authors.affiliation: Department of Biological Science, Florida State University, Tallahassee, FL 32306-4370, USA.
authors.forename: Wu-Min
authors.initials: WM
authors.lastname: Deng
mesh_terms: D000818:Animals; D002453:Cell Cycle / Q000502:physiology*; D002454:Cell Differentiation / Q000502:physiology*; D004330:Drosophila; D029721:Drosophila Proteins / Q000096:biosynthesis / Q000235:genetics / Q000378:metabolism*; D005260:Female; D018398:Homeodomain Proteins / Q000096:biosynthesis / Q000235:genetics; D009687:Nuclear Proteins / Q000096:biosynthesis / Q000235:genetics; D009866:Oogenesis / Q000502:physiology*; D006080:Ovarian Follicle / Q000166:cytology / Q000378:metabolism*; D034622:RNA Interference; D051880:Receptors, Notch / Q000235:genetics / Q000378:metabolism*; D014157:Transcription Factors / Q000096:biosynthesis / Q000235:genetics
chemical_list: D029721:Drosophila Proteins; D018398:Homeodomain Proteins; C497759:N protein, Drosophila; D009687:Nuclear Proteins; D051880:Receptors, Notch; D014157:Transcription Factors; C066381:ct protein, Drosophila