Abstract:
:Plasmid encoded replication initiation (Rep) proteins recruit host helicases to plasmid replication origins. Previously, we showed that RepD recruits directionally the PcrA helicase to the pC221 oriD, remains associated with it, and increases its processivity during plasmid unwinding. Here we show that RepD forms a complex extending upstream and downstream of the core oriD. Binding of RepD causes remodelling of a region upstream from the core oriD forming a 'landing pad' for the PcrA. PcrA is recruited by this extended RepD-DNA complex via an interaction with RepD at this upstream site. PcrA appears to have weak affinity for this region even in the absence of RepD. Upon binding of ADPNP (non-hydrolysable analogue of ATP), by PcrA, a conformational rearrangement of the RepD-PcrA-ATP initiation complex confines it strictly within the boundaries of the core oriD. We conclude that RepD-mediated recruitment of PcrA at oriD is a three step process. First, an extended RepD-oriD complex includes a region upstream from the core oriD; second, the PcrA is recruited to this upstream region and thirdly upon ATP-binding PcrA relocates within the core oriD.
journal_name
Nucleic Acids Resjournal_title
Nucleic acids researchauthors
Machón C,Lynch GP,Thomson NH,Scott DJ,Thomas CD,Soultanas Pdoi
10.1093/nar/gkp1153subject
Has Abstractpub_date
2010-04-01 00:00:00pages
1874-88issue
6eissn
0305-1048issn
1362-4962pii
gkp1153journal_volume
38pub_type
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journal_title:Nucleic acids research
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journal_title:Nucleic acids research
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