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The Lee Lab at Vanderbilt

Drosophila genome-scale screen for PAN GU kinase substrates identifies Mat89Bb as a cell cycle regulator.

AUTHORS

Lee* LALaura A , Lee* E Ethan , Anderson MA Michael A , Vardy L Leah , Tahinci E Emilios , Ali SM Siraj M , Kashevsky H Helena , Benasutti M Matt , Kirschner MW Marc W , Orr-Weaver TL Terry L . Developmental Cell. 2005 3 ; 8(3). 435-42

ABSTRACT

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.