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Vol. 15, Issue 3, 1425-1435, March 2004

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Ago1 and Dcr1, Two Core Components of the RNA Interference Pathway, Functionally Diverge from Rdp1 in Regulating Cell Cycle Events in Schizosaccharomyces pombe

Jon B. Carmichael ^*, Patrick Provost , Karl Ekwall , and Tom C. Hobman ^* 

^* Department of Cell Biology, University of Alberta, Edmonton, Alberta, Canada T6G 2H7; Centre de Recherche en Rhumatologie et Immunologie, Centre de Recherche du CHUL, Ste-Foy, Quebec, Canada G1V 4G2; and Department of Natural Sciences, Karolinska Institute, Södertörn, University College, S-141 89 Huddinge, Sweden

Submitted June 24, 2003; Revised October 10, 2003; Accepted November 10, 2003
Monitoring Editor: Suzanne Pfeffer

In the fission yeast Schizosaccharomyces pombe, three genes that function in the RNA interference (RNAi) pathway, ago1⁺, dcr1⁺, and rdp1⁺, have recently been shown to be important for timely formation of heterochromatin and accurate chromosome segregation. In the present study, we present evidence that null mutants for ago1⁺ and dcr1⁺ but not rdp1⁺, exhibit abnormal cytokinesis, cell cycle arrest deficiencies, and mating defects. Subsequent analyses showed that ago1⁺ and dcr1⁺ are required for regulated hyperphosphorylation of Cdc2 when encountering genotoxic insults. Because rdp1⁺ is dispensable for this process, the functions of ago1⁺ and dcr1⁺ in this pathway are presumably independent of their roles in RNAi-mediated heterochromatin formation and chromosome segregation. This was further supported by the finding that ago1⁺ is a multicopy suppressor of the S-M checkpoint deficiency and cytokinesis defects associated with loss of Dcr1 function, but not for the chromosome segregation defects of this mutant. Accordingly, we conclude that Dcr1-dependent production of small interfering RNAs is not required for enactment and/or maintenance of certain cell cycle checkpoints and that Ago1 and Dcr1 functionally diverge from Rdp1 to control cell cycle events in fission yeast. Finally, exogenous expression of hGERp95/EIF2C2/hAgo2, a human Ago1 homolog implicated in posttranscriptional gene silencing, compensated for the loss of ago1⁺ function in S. pombe. This suggests that PPD proteins may also be important for regulation of cell cycle events in higher eukaryotes.

Corresponding author. E-mail address: tom.hobman{at}ualberta.ca.

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