We have compared the periodic fluctuation of mRNAs encoded by CDC6, a cell cycle gene controlling initiation of DNA replication, and CLN1, a G1 cyclin gene expressed at late G1. The maxima of CDC6 mRNA precede those of CLN1 mRNA by about 15 min in cells synchronized by release from pheromone arrest or from a cdc15 thermal arrest. CDC6 mRNA accumulates in cdc15-arrested telophase cells, decays around cell separation, and reappears during telophase and nuclear division of the next cycle. CDC6 transcription at late mitosis is not affected by the pheromone signaling pathway. The CDC6 mRNA fluctuation pattern is imposed to a CLN1-derived reporter gene if fused to the CDC6 promoter. The CDC6 gene was expressed in Escherichia coli as a glutathione-S-transferase fusion protein and purified by affinity chromatography. The Cdc6 protein binds rATP and rGTP upon UV cross-linking and catalyzes the DNA-independent hydrolysis of purine nucleoside triphosphates, but does not appear to interact directly with DNA. The Cdc6 protein may control the ATP-dependent initiation of replication by conferring ATPase activity to an origin-recognizing complex.

• PMID: 8083240
• PubMed

Reference Type

Journal Article | Research Support, Non-U.S. Gov't

Authors

Zwerschke W, Rottjakob HW, Küntzel H

Primary Lit For

Additional Lit For

Review For