Positive and negative cooperativity of modularly assembled zinc fingers

Biochem Biophys Res Commun. 2009 Sep 25;387(3):440-3. doi: 10.1016/j.bbrc.2009.07.059. Epub 2009 Jul 16.

Abstract

One simple and widespread method to create engineered zinc fingers targeting the desired DNA sequences is to modularly assemble multiple finger modules pre-selected to recognize each DNA triplet. However, it has become known that a sufficient DNA binding affinity is not always obtained. In order to create successful zinc finger proteins, it is important to understand the context-dependent contribution of each finger module to the DNA binding ability of the assembled zinc finger proteins. Here, we have created finger-deletion mutants of zinc finger proteins and examined the DNA bindings of these zinc fingers to clarify the contributions of each finger module. Our results indicate that not only a positive cooperativity but also a context-dependent reduction in the DNA binding activity can be induced by assembling zinc finger modules.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Base Sequence
  • DNA / genetics
  • DNA / metabolism*
  • DNA-Binding Proteins / genetics
  • DNA-Binding Proteins / metabolism*
  • E-Box Elements*
  • Intracellular Signaling Peptides and Proteins / genetics
  • Mice
  • Period Circadian Proteins
  • Promoter Regions, Genetic
  • Sequence Deletion
  • Transcription, Genetic*
  • Zinc Fingers / genetics
  • Zinc Fingers / physiology*

Substances

  • DNA-Binding Proteins
  • Intracellular Signaling Peptides and Proteins
  • Per1 protein, mouse
  • Period Circadian Proteins
  • DNA