title: A role for homologous recombination proteins in cell cycle regulation.
abstract: Eukaryotic cells respond to DNA breaks, especially double-stranded breaks (DSBs), by activating the DNA damage response (DDR), which encompasses DNA repair and cell cycle checkpoint signaling. The DNA damage signal is transmitted to the checkpoint machinery by a network of specialized DNA damage-recognizing and signal-transducing molecules. However, recent evidence suggests that DNA repair proteins themselves may also directly contribute to the checkpoint control. Here, we investigated the role of homologous recombination (HR) proteins in normal cell cycle regulation in the absence of exogenous DNA damage. For this purpose, we used Chinese Hamster Ovary (CHO) cells expressing the Fluorescent ubiquitination-based cell cycle indicators (Fucci). Systematic siRNA-mediated knockdown of HR genes in these cells demonstrated that the lack of several of these factors alters cell cycle distribution, albeit differentially. The knock-down of MDC1, Rad51 and Brca1 caused the cells to arrest in the G2 phase, suggesting that they may be required for the G2/M transition. In contrast, inhibition of the other HR factors, including several Rad51 paralogs and Rad50, led to the arrest in the G1/G0 phase. Moreover, reduced expression of Rad51B, Rad51C, CtIP and Rad50 induced entry into a quiescent G0-like phase. In conclusion, the lack of many HR factors may lead to cell cycle checkpoint activation, even in the absence of exogenous DNA damage, indicating that these proteins may play an essential role both in DNA repair and checkpoint signaling.
authors.affiliation: a Institute of Biotechnology ; University of Lausanne ; and Center for Biotechnology UNIL-EPFL ; Lausanne , Switzerland.
authors.forename: Kaja
authors.initials: K
authors.lastname: Kostyrko
authors.forename: Sandra
authors.initials: S
authors.lastname: Bosshard
authors.forename: Zuzanna
authors.initials: Z
authors.lastname: Urban
authors.forename: Nicolas
authors.initials: N
authors.lastname: Mermod
mesh_terms: D000818:Animals; D016466:CHO Cells; D002453:Cell Cycle / Q000502:physiology*; D018797:Cell Cycle Proteins / Q000502:physiology*; D006224:Cricetinae; D003412:Cricetulus; D059765:Homologous Recombination / Q000502:physiology*; D006801:Humans; D011994:Recombinant Proteins
chemical_list: D018797:Cell Cycle Proteins; D011994:Recombinant Proteins
keywords: CHO cells; Fucci; cell cycle; homologous recombination repair; siRNA