Identification of Hedgehog pathway components by RNAi in Drosophila cultured cells.
Classical genetic screens can be limited by the selectivity of mutational targeting, the complexities of anatomically based phenotypic analysis, or difficulties in subsequent gene identification. Focusing on signaling response to the secreted morphogen Hedgehog (Hh), we used RNA interference (RNAi) and a quantitative cultured cell assay to systematically screen functional roles of all kinases and phosphatases, and subsequently 43% of predicted Drosophila genes. Two gene products reported to function in Wingless (Wg) signaling were identified as Hh pathway components: a cell surface protein (Dally-like protein) required for Hh signal reception, and casein kinase 1alpha, a candidate tumor suppressor that regulates basal activities of both Hh and Wg pathways. This type of cultured cell-based functional genomics approach may be useful in the systematic analysis of other biological processes.
Lum
Lawrence
L
Department of Molecular Biology and Genetics, Howard Hughes Medical Institute, Johns Hopkins University School of Medicine, Baltimore, MD 21205, USA.
Yao
Shenqin
S
Mozer
Brian
B
Rovescalli
Alessandra
A
Von Kessler
Doris
D
Nirenberg
Marshall
M
Beachy
Philip A
PA
D000818:Animals; D047388:Casein Kinases; D002478:Cells, Cultured; D019295:Computational Biology; D000013:Congenital Abnormalities / Q000235:genetics; D017868:Cyclic AMP-Dependent Protein Kinases / Q000235:genetics / Q000378:metabolism; D004330:Drosophila / Q000196:embryology / Q000235:genetics* / Q000378:metabolism; D029721:Drosophila Proteins / Q000235:genetics* / Q000378:metabolism; D004625:Embryo, Nonmammalian / Q000378:metabolism; D005786:Gene Expression Regulation; D017344:Genes, Insect; D016678:Genome; D023281:Genomics; D053823:Hedgehog Proteins; D009369:Neoplasms / Q000235:genetics; D011494:Protein Kinases / Q000235:genetics / Q000378:metabolism; D017346:Protein Serine-Threonine Kinases / Q000235:genetics / Q000378:metabolism; D011509:Proteoglycans / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D011518:Proto-Oncogene Proteins / Q000235:genetics / Q000378:metabolism; D034622:RNA Interference; D012330:RNA, Double-Stranded / Q000235:genetics; D015398:Signal Transduction; D014162:Transfection; D051155:Wnt1 Protein
D029721:Drosophila Proteins; D053823:Hedgehog Proteins; D011509:Proteoglycans; D011518:Proto-Oncogene Proteins; D012330:RNA, Double-Stranded; D051155:Wnt1 Protein; C420997:dlp protein, Drosophila; C074622:wg protein, Drosophila; C077245:hh protein, Drosophila; D011494:Protein Kinases; C085770:fu protein, Drosophila; D047388:Casein Kinases; D017346:Protein Serine-Threonine Kinases; D017868:Cyclic AMP-Dependent Protein Kinases