Protein fragment complementation in M.HhaI DNA methyltransferase

Wonchae Choe, Srinivasan Chandrasegaran, Marc Ostermeier

Research output: Contribution to journalArticlepeer-review

Abstract

The 5mC DNA methyltransferase M.HhaI can be split into two individually inactive N- and C-terminal fragments that together can form an active enzyme in vivo capable of efficiently methylating DNA. This active fragment pair was identified by creating libraries of M.HhaI gene fragment pairs and then selecting for the pairs that code for an active 5mC methyltransferase. The site of bisection for successful protein fragment complementation in M.HhaI was in the variable region near the target recognition domain between motif VIII and TRD. This same region is the location of bifurcation in the naturally split 5mC methyltransferase M.AquI, the location for circular permutation in M.BssHII, and the location for previously engineered split versions of M.BspRI.

Original languageEnglish (US)
Pages (from-to)1233-1240
Number of pages8
JournalBiochemical and Biophysical Research Communications
Volume334
Issue number4
DOIs
StatePublished - Sep 9 2005

Keywords

  • DNA methylation
  • DNA methyltransferase
  • Incremental truncation
  • Leucine zipper
  • M.AquI
  • M.BspRI
  • M.BssHII
  • M.HhaI
  • Protein engineering
  • Protein fragment complementation

ASJC Scopus subject areas

  • Biophysics
  • Biochemistry
  • Molecular Biology
  • Cell Biology

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