Single-molecule FRET analysis of DNA binding and bending by yeast HMGB protein Nhp6A

Julie E. Coats, Yuyen Lin, Emily Rueter, L James Maher III, Ivan Rasnik

Research output: Contribution to journalArticle

15 Citations (Scopus)

Abstract

High-mobility group B (HMGB) proteins bind duplex DNA without sequence specificity, facilitating the formation of compact nucleoprotein structures by increasing the apparent flexibility of DNA through the introduction of DNA kinks. It has remained unclear whether HMGB binding and DNA kinking are simultaneous and whether the induced kink is rigid (static) or flexible. The detailed molecular mechanism of HMGB-induced DNA 'softening' is explored here by single-molecule fluorescence resonance energy transfer studies of single yeast Nhp6A (yNhp6A) proteins binding to short DNA duplexes. We show that the local effect of yNhp6A protein binding to DNA is consistent with formation of a single static kink that is short lived (lifetimes of a few seconds) under physiological buffer conditions. Within the time resolution of our experiments, this static kink occurs at the instant the protein binds to the DNA, and the DNA straightens at the instant the protein dissociates from the DNA. Our observations support a model in which HMGB proteins soften DNA through random dynamic binding and dissociation, accompanied by DNA kinking and straightening, respectively.

Original languageEnglish (US)
Pages (from-to)1372-1381
Number of pages10
JournalNucleic Acids Research
Volume41
Issue number2
DOIs
StatePublished - Jan 2013

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High Mobility Group Proteins
Fungal Proteins
DNA
Protein Binding
Fluorescence Resonance Energy Transfer
Nucleoproteins
Buffers
Proteins

ASJC Scopus subject areas

  • Genetics

Cite this

Single-molecule FRET analysis of DNA binding and bending by yeast HMGB protein Nhp6A. / Coats, Julie E.; Lin, Yuyen; Rueter, Emily; Maher III, L James; Rasnik, Ivan.

In: Nucleic Acids Research, Vol. 41, No. 2, 01.2013, p. 1372-1381.

Research output: Contribution to journalArticle

Coats, Julie E. ; Lin, Yuyen ; Rueter, Emily ; Maher III, L James ; Rasnik, Ivan. / Single-molecule FRET analysis of DNA binding and bending by yeast HMGB protein Nhp6A. In: Nucleic Acids Research. 2013 ; Vol. 41, No. 2. pp. 1372-1381.
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