Drosophila genome-scale screen for PAN GU kinase substrates identifies Mat89Bb as a cell cycle regulator.
Although traditional organism-based mutational analysis is powerful in identifying genes involved in specific biological processes, limitations include incomplete coverage and time required for gene identification. Biochemical screens using cell transfection or yeast two-hybrid methods are rapid, but they are limited by cDNA library quality. The recent establishment of "uni-gene sets" has made it feasible to biochemically screen an organism's entire genome. Radiolabeled protein pools prepared from the Drosophila Gene Collection were used in a Drosophila in vitro expression cloning ("DIVEC") screen for substrates of PAN GU kinase, which is crucial for S-M embryonic cell cycles. Ablation of one identified substrate, Mat89Bb, by RNAi produces a polyploid phenotype similar to that of pan gu mutants. Xenopus embryos injected with Mat89Bb morpholinos arrest with polyploid nuclei, and Mat89Bb RNAi in HeLa cells gives rise to multinucleated cells. Thus, Mat89Bb plays an evolutionarily conserved role as a crucial regulator of both cell cycle and development.
Lee
Laura A
LA
Whitehead Institute, Cambridge, Massachusetts 02142, USA.
Lee
Ethan
E
Anderson
Michael A
MA
Vardy
Leah
L
Tahinci
Emilios
E
Ali
Siraj M
SM
Kashevsky
Helena
H
Benasutti
Matt
M
Kirschner
Marc W
MW
Orr-Weaver
Terry L
TL
D000818:Animals; D002453:Cell Cycle / Q000502:physiology*; D003001:Cloning, Molecular; D004330:Drosophila / Q000196:embryology / Q000201:enzymology; D029721:Drosophila Proteins / Q000235:genetics / Q000378:metabolism*; D004625:Embryo, Nonmammalian / Q000166:cytology / Q000201:enzymology*; D047108:Embryonic Development / Q000502:physiology*; D016678:Genome; D006367:HeLa Cells; D006801:Humans; D017346:Protein Serine-Threonine Kinases / Q000235:genetics / Q000378:metabolism*; D013379:Substrate Specificity / Q000235:genetics; D014981:Xenopus / Q000196:embryology / Q000378:metabolism
D029721:Drosophila Proteins; C417221:png protein, Drosophila; D017346:Protein Serine-Threonine Kinases