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Protocol for cell image-based spatiotemporal proteomics in budding yeast.

Athanasios Litsios1, Myra Paz David Masinas1, Helena Friesen1

  • 1Donnelly Centre for Cellular and Biomolecular Research, University of Toronto, Toronto, ON M5S 3E1, Canada.

STAR Protocols
|January 17, 2025
PubMed
Summary
This summary is machine-generated.

This study introduces a live-cell imaging protocol to track protein changes throughout the budding yeast cell division cycle. This method enables spatiotemporal proteome analysis for studying cell cycle transitions.

Keywords:
Cell BiologyMicroscopySingle CellSystems biology

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Area of Science:

  • Cell Biology
  • Biochemistry
  • Microscopy

Background:

  • The cell division cycle in eukaryotes involves dynamic changes in protein localization and abundance.
  • These protein dynamics are crucial for regulating key transitions during cell division.

Purpose of the Study:

  • To present a detailed protocol for spatiotemporal proteome analysis during the budding yeast cell division cycle.
  • To enable researchers to visualize and quantify protein behavior in live cells.

Main Methods:

  • Development of a live-cell imaging protocol.
  • Strain construction and cultivation of budding yeast.
  • Microscopy and subsequent image analysis for proteomic data.

Main Results:

  • A comprehensive protocol for spatiotemporal proteome analysis is described.
  • The protocol allows for the study of protein dynamics during the cell cycle.
  • Adaptability of the protocol for various genetic and environmental conditions is highlighted.

Conclusions:

  • The presented protocol facilitates in-depth analysis of the proteome during the cell division cycle.
  • This method provides a valuable tool for understanding cell cycle regulation in budding yeast.
  • The protocol's versatility extends its application to diverse research scenarios.