This Article Full Text Full Text (PDF) Alert me when this article is cited Alert me if a correction is posted Services Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Reprints and Permissions Copyright Information Books from ASM Press MicrobeWorld Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Watanabe, K. Articles by Maki, H. Search for Related Content PubMed PubMed Citation Articles by Watanabe, K. Articles by Maki, H.

 Previous Article  |  Next Article 

Eukaryotic Cell, April 2002, p. 200-212, Vol. 1, No. 2
1535-9778/02/$04.00+0     DOI: 10.1128/EC.1.2.200-212.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.

Involvement of RAD9-Dependent Damage Checkpoint Control in Arrest of Cell Cycle, Induction of Cell Death, and Chromosome Instability Caused by Defects in Origin Recognition Complex in Saccharomyces cerevisiae

Keiichi Watanabe, Jun Morishita,^, Keiko Umezu, Katsuhiko Shirahige,^, and Hisaji Maki^*

Department of Molecular Biology, Graduate School of Biological Sciences, Nara Institute of Science and Technology (NAIST), Ikoma, Nara 630-0101, Japan

Received 9 June 2001/ Accepted 10 December 2001

Perturbation of origin firing in chromosome replication is a possible cause of spontaneous chromosome instability in multireplicon organisms. Here, we show that chromosomal abnormalities, including aneuploidy and chromosome rearrangement, were significantly increased in yeast diploid cells with defects in the origin recognition complex. The cell cycle of orc1-4/orc1-4 temperature-sensitive mutant was arrested at the G₂/M boundary, after several rounds of cell division at the restrictive temperature. However, prolonged incubation of the mutant cells at 37°C led to abrogation of G₂ arrest, and simultaneously the cells started to lose viability. A sharp increase in chromosome instability followed the abrogation of G₂ arrest. In orc1-4/orc1-4 rad9/rad9 diploid cells grown at 37°C, G₂ arrest and induction of cell death were suppressed, while chromosome instability was synergistically augmented. These findings indicated that DNA lesions caused by a defect in Orc1p function trigger the RAD9-dependent checkpoint control, which ensures genomic integrity either by stopping the cell cycle progress until lesion repair, or by inducing cell death when the lesion is not properly repaired. At semirestrictive temperatures, orc2-1/orc2-1 diploid cells demonstrated G₂ arrest and loss of cell viability, both of which require RAD9-dependent checkpoint control. However, chromosome instability was not induced in orc2-1/orc2-1 cells, even in the absence of the checkpoint control. These data suggest that once cells lose the damage checkpoint control, perturbation of origin firing can be tolerated by the cells. Furthermore, although a reduction in origin-firing capacity does not necessarily initiate chromosome instability, the Orc1p possesses a unique function, the loss of which induces instability in the chromosome.

Present address: Department of Biophysics, Graduate School of Science, Kyoto University, Sakyo-ku, Kyoto 606-8502, Japan.

Present address: Genome Science Center, RIKEN, Tsurumi-ku, Yokohama, Kanagawa 230-0045, Japan.

This article has been cited by other articles:

• Makise, M., Matsui, N., Yamairi, F., Takahashi, N., Takehara, M., Asano, T., Mizushima, T. (2008). Analysis of Origin Recognition Complex in Saccharomyces cerevisiae by Use of Degron Mutants. J Biochem 143: 455-465 [Abstract] [Full Text]  
• Ide, S., Watanabe, K., Watanabe, H., Shirahige, K., Kobayashi, T., Maki, H. (2007). Abnormality in Initiation Program of DNA Replication Is Monitored by the Highly Repetitive rRNA Gene Array on Chromosome XII in Budding Yeast. Mol. Cell. Biol. 27: 568-578 [Abstract] [Full Text]  
• Laha, S., Das, S. P., Hajra, S., Sau, S., Sinha, P. (2006). The budding yeast protein Chl1p is required to preserve genome integrity upon DNA damage in S-phase. Nucleic Acids Res 34: 5880-5891 [Abstract] [Full Text]  
• Gibson, D. G., Bell, S. P., Aparicio, O. M. (2006). Cell cycle execution point analysis of ORC function and characterization of the checkpoint response to ORC inactivation in Saccharomyces cerevisiae. GENES CELLS 11: 557-573 [Abstract] [Full Text]  
• Queralt, E., Igual, J. C. (2005). Functional Connection Between the Clb5 Cyclin, the Protein Kinase C Pathway and the Swi4 Transcription Factor in Saccharomyces cerevisiae. Genetics 171: 1485-1498 [Abstract] [Full Text]  
• Weinberger, M., Ramachandran, L., Feng, L., Sharma, K., Sun, X., Marchetti, M., Huberman, J. A., Burhans, W. C. (2005). Apoptosis in budding yeast caused by defects in initiation of DNA replication. J. Cell Sci. 118: 3543-3553 [Abstract] [Full Text]  
• Gibson, D. G., Aparicio, J. G., Hu, F., Aparicio, O. M. (2004). Diminished S-Phase Cyclin-Dependent Kinase Function Elicits Vital Rad53-Dependent Checkpoint Responses in Saccharomyces cerevisiae. Mol. Cell. Biol. 24: 10208-10222 [Abstract] [Full Text]  
• Banerjee, S., Myung, K. (2004). Increased Genome Instability and Telomere Length in the elg1-Deficient Saccharomyces cerevisiae Mutant Are Regulated by S-Phase Checkpoints. Eukaryot Cell 3: 1557-1566 [Abstract] [Full Text]  
• Myung, K., Pennaneach, V., Kats, E. S., Kolodner, R. D. (2003). Saccharomyces cerevisiae chromatin-assembly factors that act during DNA replication function in the maintenance of genome stability. Proc. Natl. Acad. Sci. USA 100: 6640-6645 [Abstract] [Full Text]  
• Yoshida, J., Umezu, K., Maki, H. (2003). Positive and Negative Roles of Homologous Recombination in the Maintenance of Genome Stability in Saccharomyces cerevisiae. Genetics 164: 31-46 [Abstract] [Full Text]