Bacterial origin recognition complexes direct assembly of higher-order DnaA oligomeric structures

Proc Natl Acad Sci U S A. 2009 Nov 3;106(44):18479-84. doi: 10.1073/pnas.0909472106. Epub 2009 Oct 15.

Abstract

Eukaryotic initiator proteins form origin recognition complexes (ORCs) that bind to replication origins during most of the cell cycle and direct assembly of prereplication complexes (pre-RCs) before the onset of S phase. In the eubacterium Escherichia coli, there is a temporally similar nucleoprotein complex comprising the initiator protein DnaA bound to three high-affinity recognition sites in the unique origin of replication, oriC. At the time of initiation, this high-affinity DnaA-oriC complex (the bacterial ORC) accumulates additional DnaA that interacts with lower-affinity sites in oriC, forming a pre-RC. In this paper, we investigate the functional role of the bacterial ORC and examine whether it mediates low-affinity DnaA-oriC interactions during pre-RC assembly. We report that E. coli ORC is essential for DnaA occupation of low-affinity sites. The assistance given by ORC is directed primarily to proximal weak sites and requires oligomerization-proficient DnaA. We propose that in bacteria, DnaA oligomers of limited length and stability emerge from single high-affinity sites and extend toward weak sites to facilitate their loading as a key stage of prokaryotic pre-RC assembly.

Publication types

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

MeSH terms

  • Bacterial Proteins / chemistry*
  • Bacterial Proteins / metabolism*
  • Base Sequence
  • Binding Sites
  • DNA-Binding Proteins / chemistry*
  • DNA-Binding Proteins / metabolism*
  • Escherichia coli / metabolism*
  • Molecular Sequence Data
  • Mutation / genetics
  • Origin Recognition Complex / metabolism*
  • Protein Binding
  • Protein Structure, Quaternary
  • Replication Origin / genetics

Substances

  • Bacterial Proteins
  • DNA-Binding Proteins
  • DnaA protein, Bacteria
  • Origin Recognition Complex