Jove
Visualize
Contact Us
JoVE
x logofacebook logolinkedin logoyoutube logo
ABOUT JoVE
OverviewLeadershipBlogJoVE Help Center
AUTHORS
Publishing ProcessEditorial BoardScope & PoliciesPeer ReviewFAQSubmit
LIBRARIANS
TestimonialsSubscriptionsAccessResourcesLibrary Advisory BoardFAQ
RESEARCH
JoVE JournalMethods CollectionsJoVE Encyclopedia of ExperimentsArchive
EDUCATION
JoVE CoreJoVE BusinessJoVE Science EducationJoVE Lab ManualFaculty Resource CenterFaculty Site
Terms & Conditions of Use
Privacy Policy
Policies

Related Experiment Videos

Chromatin assembly in yeast cell-free extracts.

M C Schultz1

  • 1Department of Biochemistry, University of Alberta, Edmonton, Alberta, T6G 2H7, Canada.michael.schultz@ualberta.ca

Methods (San Diego, Calif.)
|March 17, 1999
PubMed
Summary

Researchers developed a simple method to create chromatin assembly extracts from budding yeast. This breakthrough enables combined biochemical and genetic studies of chromatin metabolism, previously not possible in this model organism.

Related Concept Videos

You might also read

Related Articles

Articles linked to this work by shared authors, journal, and citation graph.

Sort by
Same author

TATA binding protein-associated CK2 transduces DNA damage signals to the RNA polymerase III transcriptional machinery.

Cell·2001
Same author

A review of progress towards elucidating the role of protein kinase CK2 in polymerase III transcription: regulation of the TATA binding protein.

Molecular and cellular biochemistry·1999
Same author

Histone modification governs the cell cycle regulation of a replication-independent chromatin assembly pathway in Saccharomyces cerevisiae.

Proceedings of the National Academy of Sciences of the United States of America·1999
Same author

Rapamycin induces the G0 program of transcriptional repression in yeast by interfering with the TOR signaling pathway.

Molecular and cellular biology·1998
Same author

Deposition-related sites K5/K12 in histone H4 are not required for nucleosome deposition in yeast.

Proceedings of the National Academy of Sciences of the United States of America·1998
Same author

Casein kinase II regulation of yeast TFIIIB is mediated by the TATA-binding protein.

Genes & development·1997

Area of Science:

  • Molecular Biology
  • Biochemistry
  • Genetics

Background:

  • Chromatin assembly extracts are crucial for studying DNA replication and repair.
  • A simple, effective method for preparing these extracts from budding yeast (Saccharomyces cerevisiae) was previously unavailable.
  • Budding yeast is a powerful model organism for genetic studies.

Purpose of the Study:

  • To describe a simple and detailed method for preparing chromatin assembly extracts from budding yeast.
  • To characterize the composition and function of these extracts.
  • To enable combined biochemical and genetic analyses of chromatin metabolism in yeast.

Main Methods:

  • Preparation of a crude 100,000g supernatant from frozen, disrupted budding yeast cells.
  • Assessment of chromatin assembly activity in the extract.
  • Analysis of the extract's dependence on ATP and specific histone modifications.

Main Results:

  • The prepared extract contains core histones and necessary soluble protein factors for chromatin assembly under physiological conditions.
  • Chromatin assembly in the extract is sensitive to mutations in histone H4 N-terminal tail lysine residues, correlating with in vivo acetylation.
  • The ATP-dependent reaction drives DNA supercoiling with efficiency comparable to mammalian cell extracts.

Conclusions:

  • A simple method for preparing functional chromatin assembly extracts from budding yeast has been established.
  • This system facilitates the study of chromatin metabolism using a combined biochemical and genetic approach in yeast.
  • The findings provide a new tool for understanding fundamental processes of chromatin dynamics and regulation.

Related Experiment Videos