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PMID: 9008708 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Cell cycle regulation of the replication licensing system: involvement of a Cdk-dependent inhibitor.

The Journal of cell biology ·Vol. 136 ·No. 1 ·1997-01-13 ·Pages 125-35

Mahbubani HM, Chong JP, Chevalier S, Thömmes P, Blow JJ

Abstract

The replication licensing factor (RLF) is an essential initiation factor that is involved in preventing re-replication of chromosomal DNA in a single cell cycle. In Xenopus egg extracts, it can be separated into two components: RLF-M, a complex of MCM/P1 polypeptides, and RLF-B, which is currently unpurified. In this paper we investigate variations in RLF activity throughout the cell cycle. Total RLF activity is low in metaphase, due to a lack of RLF-B activity and the presence of an RLF inhibitor. RLF-B is rapidly activated on exit from metaphase, and then declines during interphase. The RLF inhibitor present in metaphase extracts is dependent on the activity of cyclin-dependent kinases (Cdks). Affinity depletion of Cdks from metaphase extracts removed the RLF inhibitor, while Cdc2/cyclin B directly inhibited RLF activity. In metaphase extracts treated with the protein kinase inhibitor 6-dimethylaminopurine (6-DMAP), both cyclin B and the RLF inhibitor were stabilized although the extracts morphologically entered interphase. These results are consistent with studies in other organisms that invoke a key role for Cdks in preventing re-replication of DNA in a single cell cycle.

MeSH Terms
Adenine/analogs & derivatives,pharmacology Animals Cell Cycle/physiology Cell Cycle Proteins/metabolism,pharmacology,physiology Cyclin-Dependent Kinase Inhibitor p21 Cyclin-Dependent Kinases/antagonists & inhibitors,physiology Cyclins/pharmacology Cycloheximide/pharmacology DNA Replication/drug effects,physiology Enzyme Inhibitors/pharmacology Protein Synthesis Inhibitors/pharmacology Xenopus
Chemicals
Cell Cycle Proteins Cyclin-Dependent Kinase Inhibitor p21 Cyclins Enzyme Inhibitors Protein Synthesis Inhibitors N(6),N(6)-dimethyladenine Cycloheximide Cyclin-Dependent Kinases Adenine
Authors & Affiliations
5 authors, click to expand affiliations / ORCID
Mahbubani H M
Imperial Cancer Research Fund, Clare Hall Laboratories, Potters Bar, Herts, United Kingdom.
Chong J P
Chevalier S
Thömmes P
Blow J J
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Article Info
Journal
The Journal of cell biology
Abbr.
J Cell Biol
ISSN
0021-9525
DOI
10.1083/jcb.136.1.125
Published
1997-01-13
Pages
125-35
Language
English
Region
United States
NLM ID
0375356
PMCID
PMC2132454
Subset
IM
External Links
PubMed source DOI: 10.1083/jcb.136.1.125 PMC Full Text View journal details
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