A fluorescence resonance energy transfer-based probe to monitor nucleosome structure

D. Lovullo, D. Daniel, J. Yodh, D. Lohr, Neal Woodbury

Research output: Contribution to journalArticlepeer-review

18 Scopus citations

Abstract

Nucleosomes are the basic units of eukaryotic chromatin structure. By restricting factor access to regulatory DNA sequences, nucleosomes significantly impact genomic processes such as transcription, and various mechanisms to alter nucleosome structure to relieve this repression have evolved. Both nucleosomes and processes that alter them are inherently dynamic in nature. Thus, studies of dynamics will be necessary to truly understand these relief mechanisms. We describe here the characteristics of a novel fluorescence resonance energy transfer-based reporter that can clearly signal the formation of a canonical nucleosome structure and follow conformational and compositional changes in that structure, both at the ensemble-average (bulk) and at the single molecule level. Labeled nucleosomes behave conformationally and thermodynamically like typical nucleosomes; thus they are relevant reporters of nucleosome behavior. Nucleosomes and free DNA are readily distinguishable at the single-molecule level. Thus, these labeled nucleosomes are well suited to studies of dynamic changes in nucleosome structure including single-molecule dynamics.

Original languageEnglish (US)
Pages (from-to)165-172
Number of pages8
JournalAnalytical Biochemistry
Volume341
Issue number1
DOIs
StatePublished - Jun 1 2005

Keywords

  • Chromatin
  • FRET
  • Nucleosome
  • Single-molecule spectroscopy

ASJC Scopus subject areas

  • Biophysics
  • Biochemistry
  • Molecular Biology
  • Cell Biology

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