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Related Experiment Videos

Yeast Isw1p forms two separable complexes in vivo.

Jay C Vary1, Vamsi K Gangaraju, Jun Qin

  • 1Division of Basic Sciences, Fred Hutchinson Cancer Research Center, Seattle, Washington 98109, USA.

Molecular and Cellular Biology
|December 17, 2002
PubMed
Summary
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The budding yeast Isw1 protein forms two distinct chromatin remodeling complexes, Isw1a and Isw1b. These complexes exhibit unique DNA-binding and nucleosome-sliding activities, impacting gene transcription and cell growth.

Area of Science:

  • Molecular Biology
  • Biochemistry
  • Genetics

Background:

  • ATP-dependent chromatin remodeling complexes regulate cellular processes by altering chromatin structure.
  • The budding yeast Saccharomyces cerevisiae possesses two ISWI-class ATPases: Isw1p and Isw2p.
  • Previous research indicated Isw1p copurifies with other proteins.

Purpose of the Study:

  • To identify proteins associated with Isw1p.
  • To characterize the distinct in vivo complexes formed by Isw1p (Isw1a and Isw1b).
  • To investigate the functional differences between Isw1a and Isw1b in vitro and in vivo.

Main Methods:

  • Copurification and protein identification assays.
  • Biochemical assays measuring ATPase activity, DNA binding, and nucleosome binding.

Related Experiment Videos

  • In vivo studies assessing transcriptional regulation and temperature sensitivity.

    • Main Results:

    • Isw1p forms two separable complexes in vivo: Isw1a and Isw1b.
    • Both complexes display similar nucleosome-stimulated ATPase activity.
    • Isw1a and Isw1b exhibit differential DNA and nucleosome binding affinities, leading to distinct nucleosome spacing and sliding activities.
    • The two complexes have overlapping and distinct roles in gene transcriptional regulation.
    • Isw1a and Isw1b contribute differently to cell growth at high temperatures.

    Conclusions:

    • Isw1p exists as two distinct functional complexes with specialized roles.
    • Differential substrate binding underlies the distinct activities of Isw1a and Isw1b.
    • These complexes play crucial, yet distinct, roles in gene expression and cellular adaptation to thermal stress.