Quantitative proteomic analysis of yeast DNA replication proteins

Research output: Contribution to journalArticle

12 Citations (Scopus)

Abstract

Chromatin is dynamically regulated, and proteomic analysis of its composition can provide important information about chromatin functional components. Many DNA replication proteins for example bind chromatin at specific times during the cell cycle. Proteomic investigation can also be used to characterize changes in chromatin composition in response to perturbations such as DNA damage, while useful information is obtained by testing the effects on chromatin composition of mutations in chromosome stability pathways. We have successfully used the method of stable isotope labeling by amino acids in cell culture (SILAC) for quantitative proteomic analysis of normal and pathological changes to yeast chromatin. Here we describe this proteomic method for analyzing changes to Saccharomyces cerevisiae chromatin, illustrating the procedure with an analysis of the changes that occur in chromatin composition as cells progress from a G1 phase block (induced by alpha factor) into S phase (in the presence of DNA replication inhibitor hydroxyurea).

Original languageEnglish
Pages (from-to)196-202
Number of pages7
JournalMethods
Volume57
Issue number2
Early online date23 Mar 2012
DOIs
Publication statusPublished - Jun 2012

Fingerprint

DNA Replication
Proteomics
Yeast
Chromatin
Yeasts
DNA
Proteins
Chemical analysis
Isotope Labeling
Chromosomal Instability
Hydroxyurea
G1 Phase
Chromosomes
S Phase
Cell culture
Isotopes
Labeling
DNA Damage
Saccharomyces cerevisiae
Cell Cycle

Keywords

  • quantitative proteomics
  • SILAC
  • chromatin
  • chromatin preparation
  • DNA replication

Cite this

Quantitative proteomic analysis of yeast DNA replication proteins. / Kubota, Takashi; Stead, David Andrew; Hiraga, Shinichiro; ten Have, Sara; Donaldson, Anne Dunlop.

In: Methods, Vol. 57, No. 2, 06.2012, p. 196-202.

Research output: Contribution to journalArticle

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