Transcription regulation of the type II restriction-modification system AhdI

Nucleic Acids Res. 2008 Mar;36(5):1429-42. doi: 10.1093/nar/gkm1116. Epub 2008 Jan 18.

Abstract

The Restriction-modification system AhdI contains two convergent transcription units, one with genes encoding methyltransferase subunits M and S and another with genes encoding the controller (C) protein and the restriction endonuclease (R). We show that AhdI transcription is controlled by two independent regulatory loops that are well-optimized to ensure successful establishment in a naïve bacterial host. Transcription from the strong MS promoter is attenuated by methylation of an AhdI site overlapping the -10 element of the promoter. Transcription from the weak CR promoter is regulated by the C protein interaction with two DNA-binding sites. The interaction with the promoter-distal high-affinity site activates transcription, while interaction with the weaker promoter-proximal site represses it. Because of high levels of cooperativity, both C protein-binding sites are always occupied in the absence of RNA polymerase, raising a question how activated transcription is achieved. We develop a mathematical model that is in quantitative agreement with the experiment and indicates that RNA polymerase outcompetes C protein from the promoter-proximal-binding site. Such an unusual mechanism leads to a very inefficient activation of the R gene transcription, which presumably helps control the level of the endonuclease in the cell.

Publication types

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

MeSH terms

  • Binding Sites
  • Chromosome Mapping
  • DNA Footprinting
  • DNA Modification Methylases / biosynthesis
  • DNA Modification Methylases / genetics*
  • Deoxyribonucleases, Type II Site-Specific / biosynthesis
  • Deoxyribonucleases, Type II Site-Specific / genetics*
  • Gene Expression Regulation, Bacterial*
  • Models, Genetic*
  • Promoter Regions, Genetic
  • Transcription Factors / metabolism
  • Transcription, Genetic

Substances

  • Transcription Factors
  • DNA Modification Methylases
  • Deoxyribonucleases, Type II Site-Specific