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Related Experiment Videos

A proteasome cap subunit required for spindle pole body duplication in yeast

H B McDonald1, B Byers

  • 1Department of Genetics, University of Washington, Seattle 98195, USA.

The Journal of Cell Biology
|May 5, 1997
PubMed
Summary

Proteasome cap subunit Pcs1 is essential for yeast viability, regulating spindle pole body duplication. Loss of Pcs1 function causes cell cycle arrest with unsegregated DNA and enlarged spindle pole bodies.

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Saccharomyces cerevisiae MPS2 encodes a membrane protein localized at the spindle pole body and the nuclear envelope.

Molecular biology of the cell·1999

Area of Science:

  • Cell Biology
  • Molecular Biology
  • Biochemistry

Background:

  • Proteasome-mediated protein degradation regulates critical cellular processes like cell cycle progression.
  • Proteasomal 19S cap complexes control substrate entry into the 20S proteolytic core.
  • Regulation of substrate degradation may occur within these 19S cap complexes.

Purpose of the Study:

  • To characterize a novel Saccharomyces cerevisiae gene, PCS1, encoding a proteasomal 19S cap subunit.
  • To investigate the function of PCS1 in cell viability and cell cycle progression.
  • To elucidate the role of Pcs1p in spindle pole body (SPB) duplication.

Main Methods:

  • Gene characterization and sequence analysis of PCS1.
  • Construction and analysis of a temperature-sensitive pcs1 mutant strain.

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  • Electron microscopy (EM) to examine cell morphology and SPB duplication.
  • Localization studies using a functional Pcs1-green fluorescent protein (GFP) fusion.

    • Main Results:

    • PCS1 is essential for Saccharomyces cerevisiae viability.
    • A temperature-sensitive pcs1 mutant arrests in the cell cycle as large-budded cells with G2 DNA content and unsegregated DNA.
    • Mutant cells exhibit failure in spindle pole body (SPB) duplication, with enlarged SPBs.
    • Pcs1p localizes to the nucleus throughout the cell cycle.

    Conclusions:

    • Pcs1p is a crucial component of the proteasomal 19S cap complex.
    • PCS1 function is vital for cell cycle progression, specifically for SPB duplication.
    • Pcs1p likely mediates the degradation of nuclear components required for SPB duplication.