Guided nuclear exploration increases CTCF target search efficiency

Nat Chem Biol. 2020 Mar;16(3):257-266. doi: 10.1038/s41589-019-0422-3. Epub 2019 Dec 2.

Abstract

The enormous size of mammalian genomes means that for a DNA-binding protein the number of nonspecific, off-target sites vastly exceeds the number of specific, cognate sites. How mammalian DNA-binding proteins overcome this challenge to efficiently locate their target sites is not known. Here, through live-cell single-molecule tracking, we show that CCCTC-binding factor, CTCF, is repeatedly trapped in small zones that likely correspond to CTCF clusters, in a manner that is largely dependent on an internal RNA-binding region (RBRi). We develop a new theoretical model called anisotropic diffusion through transient trapping in zones to explain CTCF dynamics. Functionally, transient RBRi-mediated trapping increases the efficiency of CTCF target search by ~2.5-fold. Overall, our results suggest a 'guided' mechanism where CTCF clusters concentrate diffusing CTCF proteins near cognate binding sites, thus increasing the local ON-rate. We suggest that local guiding may allow DNA-binding proteins to more efficiently locate their target sites.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Binding Sites / physiology
  • CCCTC-Binding Factor / metabolism*
  • CCCTC-Binding Factor / physiology
  • Cell Line
  • Cell Nucleus / metabolism
  • Chromatin / metabolism
  • DNA-Binding Proteins / metabolism
  • DNA-Binding Proteins / physiology
  • Female
  • Humans
  • Male
  • Mice
  • Protein Binding / physiology
  • Repressor Proteins / metabolism
  • Single Molecule Imaging / methods*

Substances

  • CCCTC-Binding Factor
  • CTCF protein, human
  • CTCFL protein, human
  • Chromatin
  • DNA-Binding Proteins
  • Repressor Proteins