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

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.

authors

Miller DT,Grimwade JE,Betteridge T,Rozgaja T,Torgue JJ,Leonard AC

doi

10.1073/pnas.0909472106

subject

Has Abstract

pub_date

2009-11-03 00:00:00

pages

18479-84

issue

44

eissn

0027-8424

issn

1091-6490

pii

0909472106

journal_volume

106

pub_type

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