RCC1-Ran-RanGAP signal for initiation of mitosis
Conference
·
OSTI ID:28440
- Deutsches Krebsforschungszentrum, Heidelberg (Germany)
In most eukaryotic cells checkpoints ensure that DNA replication is completed before the onset of mitosis. Experimentally these controls can be overrun, e.g., by fusion of S-phase cells with mitotic cells, by adding 5 mM caffeine to the medium of S-phase cells, or by okadaic acid, resulting in premature condensation of the chromosomes and in their {open_quote}pulverized{close_quote} appearance under the microscope. One of the earliest genes involved in these controls is RCC1, the {open_quote}regulator of chromosome condensation{close_quote}. It has been identified by T. Nishimoto in a baby hamster kidney cell line BHK21 as a gene whose temperature sensitive mutation induces mitotic events independently of completed DNA replication. When mutant cells are arrested in S-phase by inhibitors of DNA synthesis, a shift to the nonpermissive temperature causes loss of the mutated RCC1 gene product and results in premature activation of the p34{sup cdc2} kinase and consequently induction of mitotic events. This requires protein synthesis, but occurs in the presence of inhibitors of transcription. Thus, active RCC1 is required to prevent induction of mitosis before DNA replication is complete, and it displays its activity upstream of the cdc2 kinase. Homologous genes have been found in Xenopus, Drosophila, Saccharomyces cerevisiae, and Saccharomyces pombe. In an investigation of putative human centromere proteins we have been able to purify RCC1 gene product. We could demonstrate its biochemical activity, isolate its immediate target, and enrich a third element of the signal chain.
- OSTI ID:
- 28440
- Report Number(s):
- CONF-9210475--Cond.
- Country of Publication:
- United States
- Language:
- English
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