The cyclin A centrosomal localization sequence recruits MCM5 and Orc1 to regulate centrosome reduplication

J Cell Sci. 2010 Aug 15;123(Pt 16):2743-9. doi: 10.1242/jcs.073098. Epub 2010 Jul 27.

Abstract

Centrosomes are the major microtubule-organizing centers in animal cells and regulate formation of a bipolar mitotic spindle. Aberrant centrosome number causes chromosome mis-segregation, and has been implicated in genomic instability and tumor development. Previous studies have demonstrated a role for the DNA replication factors MCM5 and Orc1 in preventing centrosome reduplication. Cyclin A-Cdk2 localizes on centrosomes by means of a modular centrosomal localization sequence (CLS) that is distinct from that of cyclin E. Here, we show that cyclin A interacts with both MCM5 and Orc1 in a CLS-dependent but Cdk-independent manner. Although the MRAIL hydrophobic patch is contained within the cyclin A CLS, binding of both MCM5 and Orc1 to cyclin A does not require a wild-type hydrophobic patch. The same domain in MCM5 that mediates interaction with cyclin E also binds cyclin A, resulting in centrosomal localization of MCM5. Finally, unlike its function in DNA synthesis, MCM5-mediated inhibition of centrosome reduplication in S-phase-arrested CHO cells does not require binding to other MCM family members. These results suggest that cyclins E and A sequentially prevent centrosome reduplication throughout interphase by recruitment of DNA replication factors such as MCM5 and Orc1.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • CHO Cells
  • Cell Cycle Proteins / chemistry
  • Cell Cycle Proteins / genetics
  • Cell Cycle Proteins / metabolism
  • Cell Cycle Proteins / physiology*
  • Centrosome / metabolism
  • Centrosome / physiology*
  • Cricetinae
  • Cricetulus
  • Cyclin A / genetics
  • Cyclin A / metabolism
  • Cyclin A / physiology*
  • Fluorescent Antibody Technique
  • HeLa Cells
  • Humans
  • Immunoprecipitation
  • Microtubule-Organizing Center / metabolism
  • Origin Recognition Complex / genetics
  • Origin Recognition Complex / metabolism
  • Origin Recognition Complex / physiology*
  • Transfection

Substances

  • Cell Cycle Proteins
  • Cyclin A
  • MCM5 protein, human
  • ORC1 protein, human
  • Origin Recognition Complex