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Interaction trap experiment with CDC6.

P B Zavec1, A Comino, P Watt

  • 1National Institute of Chemistry, Ljubljana, Slovenia.

Pflugers Archiv : European Journal of Physiology
|February 1, 2000
PubMed
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Researchers identified proteins interacting with the essential yeast gene CDC6, revealing roles in cell wall synthesis and potential carbon metabolism. Some interactions suggest involvement in cell cycle regulation and retrotransposition.

Area of Science:

  • Molecular Biology
  • Yeast Genetics
  • Cell Cycle Regulation

Background:

  • The gene CDC6 is crucial for Saccharomyces cerevisiae viability.
  • Despite its known importance, the precise biochemical function of the Cdc6 protein in the cell cycle is not well understood.
  • Understanding Cdc6 protein interactions is key to elucidating its role in cellular processes.

Purpose of the Study:

  • To identify proteins that specifically interact with Cdc6 using an interaction trap experiment.
  • To investigate the functional implications of these interactions for Cdc6's role in the cell cycle and other cellular processes.

Main Methods:

  • Employed the interaction trap (two-hybrid) system to screen for Cdc6-interacting proteins.
  • Detected interactions by measuring beta-galactosidase activity.

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  • Analyzed interacting gene sequences (ECM11, YNL201, YOR279) and assessed their effect on a cdc6-1 mutation.

    • Main Results:

    • Identified four interacting partners: ECM11 (cell wall synthesis), YNL201 (potential carbon metabolism), YOR279 (unknown function), and the Ty1 retrotransposition element.
    • Strongest interactions with Cdc6 were observed for ECM11 and YNL201, with weaker interactions for YOR279 and Ty1A.
    • Ty1A element showed the strongest suppression of the cdc6-1 mutation, followed by ECM11 and YNL201; YOR279 did not suppress the mutation.

    Conclusions:

    • Cdc6 protein interacts with proteins involved in diverse cellular functions, including cell wall synthesis and potentially carbon metabolism.
    • The interaction with Ty1 suggests a role for Cdc6 in regulating retrotransposition.
    • These findings provide new insights into the multifaceted functions of Cdc6 beyond its established role in DNA replication.