High Free-Energy Barrier of 1D Diffusion Along DNA by Architectural DNA-Binding Proteins

Kiyoto Kamagata, Eriko Mano, Kana Ouchi, Saori Kanbayashi, Reid C. Johnson

Research output: Contribution to journalArticlepeer-review

26 Citations (Scopus)


Architectural DNA-binding proteins function to regulate diverse DNA reactions and have the defining property of significantly changing DNA conformation. Although the 1D movement along DNA by other types of DNA-binding proteins has been visualized, the mobility of architectural DNA-binding proteins on DNA remains unknown. Here, we applied single-molecule fluorescence imaging on arrays of extended DNA molecules to probe the binding dynamics of three structurally distinct architectural DNA-binding proteins: Nhp6A, HU, and Fis. Each of these proteins was observed to move along DNA, and the salt concentration independence of the 1D diffusion implies sliding with continuous contact to DNA. Nhp6A and HU exhibit a single sliding mode, whereas Fis exhibits two sliding modes. Based on comparison of the diffusion coefficients and sizes of many DNA binding proteins, the architectural proteins are categorized into a new group distinguished by an unusually high free-energy barrier for 1D diffusion. The higher free-energy barrier for 1D diffusion by architectural proteins can be attributed to the large DNA conformational changes that accompany binding and impede rotation-coupled movement along the DNA grooves.

Original languageEnglish
Pages (from-to)655-667
Number of pages13
JournalJournal of Molecular Biology
Issue number5
Publication statusPublished - 2018 Mar 2


  • DNA conformational change
  • HMGB chromatin protein
  • bacterial nucleoid protein
  • protein–DNA sliding dynamics
  • single-molecule fluorescence microscopy


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