Cyclin A promotes S-phase entry via interaction with the replication licensing factor Mcm7.

Publication Type:

Journal Article

Source:

Molecular and cellular biology, Volume 31, Issue 2, p.248-55 (2011)

Keywords:

2011, Amino Acid Sequence, Animals, Base Sequence, Basic Sciences Division, Cell Cycle Proteins, Cell Line, Tumor, Cyclin A, DNA Replication, DNA-Binding Proteins, Humans, MICE, Molecular Sequence Data, Mutation, NIH 3T3 Cells, Nuclear Proteins, RNA Interference, S Phase, Two-Hybrid System Techniques

Abstract:

Cyclin A is known to promote S-phase entry in mammals, but its critical targets in this process have not been defined. We derived a novel human cyclin A mutant (CycA-C1), which can activate cyclin-dependent kinase but cannot promote S-phase entry, and isolated replication licensing factor Mcm7 as a factor that interacts with the wild-type cyclin A but not with the mutant. We demonstrated that human cyclin A and Mcm7 interact in the chromatin fraction. To address the physiological significance of the cyclin A-Mcm7 interaction, we isolated an Mcm7 mutant (Mcm7-3) that is capable of association with CycA-C1 and found that it can also suppress the deficiency of CycA-C1 in promoting S-phase entry. Finally, RNA interference experiments showed that the CycA-C1 mutant is defective for the endogenous cyclin A function in S-phase entry and that this defect can be suppressed by the Mcm7-3 mutant. Our findings demonstrate that interaction with Mcm7 is essential for the function of cyclin A in promoting S-phase entry.