Analysis of chromatin structure by in vivo formaldehyde cross-linking

Valerio Orlando, Helen Strutt, Renato Paro

Research output: Contribution to journalArticle

488 Citations (Scopus)

Abstract

Recent advances leave no doubt that higher order chromatin structures play a fundamental role in many developmentally important mechanisms of gene regulation. In particular analyses in genetic model systems like yeast and Drosophila uncovered novel proteins that are involved in the regulation of chromatin structures. Many of these proteins do not bind directly to DNA but interact in large multimeric complexes. To identify the DNA elements regulated by these multiprotein complexes, alternative approaches to the standard methods of DNA-protein analysis had to be devised. Here we present a method that preserves the architecture of the higher order chromatin structures by crosslinking cells in vivo with formaldehyde. An immunoprecipitation strategy is then used to identify the DNA targets of chromosomal proteins of interest. This method can be applied to study the distribution of proteins at high resolution over extended chromosomal regions.

Original languageEnglish
Pages (from-to)205-214
Number of pages10
JournalMethods in Enzymology
Volume11
Issue number2
DOIs
Publication statusPublished - Feb 1997

Fingerprint

Formaldehyde
Chromatin
DNA
Proteins
Multiprotein Complexes
Genetic Models
Immunoprecipitation
Gene expression
Crosslinking
Yeast
Drosophila
Yeasts
Genes

ASJC Scopus subject areas

  • Molecular Biology
  • Biochemistry, Genetics and Molecular Biology(all)

Cite this

Analysis of chromatin structure by in vivo formaldehyde cross-linking. / Orlando, Valerio; Strutt, Helen; Paro, Renato.

In: Methods in Enzymology, Vol. 11, No. 2, 02.1997, p. 205-214.

Research output: Contribution to journalArticle

Orlando, Valerio ; Strutt, Helen ; Paro, Renato. / Analysis of chromatin structure by in vivo formaldehyde cross-linking. In: Methods in Enzymology. 1997 ; Vol. 11, No. 2. pp. 205-214.
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