High-resolution mapping of architectural DNA binding protein facilitation of a DNA repression loop in Escherichia coli

Nicole A. Becker, L James Maher III

Research output: Contribution to journalArticle

6 Citations (Scopus)

Abstract

Double-stranded DNA is a locally inflexible polymer that resists bending and twisting over hundreds of base pairs. Despite this, tight DNA bending is biologically important for DNA packaging in eukaryotic chromatin and tight DNA looping is important for gene repression in prokaryotes. We and others have previously shown that sequence nonspecific DNA kinking proteins, such as Escherichia coli heat unstable and Saccharomyces cerevisiae non-histone chromosomal protein 6A (Nhp6A), facilitate lac repressor (LacI) repression loops in E. coli. It has been unknown if this facilitation involves direct protein binding to the tightly bent DNA loop or an indirect effect promoting global negative supercoiling of DNA. Here we adapt two high-resolution in vivo protein-mapping techniques to demonstrate direct binding of the heterologous Nhp6A protein at a LacI repression loop in living E. coli cells.

Original languageEnglish (US)
Pages (from-to)7177-7182
Number of pages6
JournalProceedings of the National Academy of Sciences of the United States of America
Volume112
Issue number23
DOIs
StatePublished - Jun 9 2015

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DNA-Binding Proteins
Escherichia coli
Non Histone Chromosomal Proteins
DNA
Lac Repressors
DNA Packaging
Saccharomyces cerevisiae Proteins
Proteins
Protein Binding
Base Pairing
Chromatin
Polymers
Hot Temperature
Genes

Keywords

  • Architectural protein
  • DNA looping
  • E. coli
  • Lac
  • Nhp6A

ASJC Scopus subject areas

  • General

Cite this

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