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The identification of Pcl1-interacting proteins that genetically interact with Cla4 may indicate a link between G1

Megan E Keniry1, Hilary A Kemp, David M Rivers

  • 1Department of Biology and Institute of Molecular Biology, University of Oregon, Eugene, Oregon 97403-1229, USA.

Genetics
|April 15, 2004
PubMed

Insights

Budding yeast

Area of Science:

  • Cell Biology
  • Molecular Biology
  • Genetics

Background:

  • Cla4 and Ste20 are p21-activated kinases crucial for budding yeast morphogenesis.
  • While individually distinct, their combined loss is lethal, indicating functional overlap.
  • Both kinases have been recently linked to mitotic exit regulation.

Purpose of the Study:

  • To investigate the roles of Cla4 and its interacting proteins in cellular processes.
  • To identify novel regulators of mitotic exit in budding yeast.
  • To elucidate the functional relationship between Pho85, Pcl1/Pcl2, and Cla4 in cell division.

Main Methods:

  • Genetic interaction studies, including lethal mutation screening in a cla4 Delta strain.
  • Two-hybrid screening to identify Pcl1-interacting proteins.
  • GST pull-down assays and in vitro phosphorylation experiments.
  • Analysis of Cdc14 localization in mutant strains.

Main Results:

  • A genetic interaction was identified between PHO85 and CLA4.
  • Pho85-coupled G(1) cyclins Pcl1 and Pcl2 were shown to contribute to Pho85's role.
  • Ncp1, Hms1, and Epa1 were identified as Pcl1 interactors and substrates.
  • NCP1, HMS1, and EPA1 also genetically interact with CLA4.
  • Hms1, Ncp1, and Pho85 were implicated in mitotic exit, evidenced by Cdc14 mislocalization.

Conclusions:

  • The G(1) cyclin-dependent kinase complex Pcl1.Pho85 plays a role in regulating mitotic exit.
  • Ncp1, Hms1, and Epa1 are novel components of the mitotic exit pathway, interacting with both Pcl1.Pho85 and Cla4.
  • A model is proposed where Pcl1.Pho85 regulates mitotic exit machinery, potentially integrating with Cla4 pathways.

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