Abstract:
:Double-stranded DNA break repair by homologous recombination is initiated by resection of free DNA ends to produce a 3'-ssDNA overhang. In bacteria, this reaction is catalyzed by helicase-nuclease complexes such as AddAB in a manner regulated by specific recombination hotspot sequences called Crossover hotspot instigator (Chi). We have used magnetic tweezers to investigate the dynamics of AddAB translocation and hotspot scanning during double-stranded DNA break resection. AddAB was prone to stochastic pausing due to transient recognition of Chi-like sequences, unveiling an antagonistic relationship between DNA translocation and sequence-specific DNA recognition. Pauses at bona fide Chi sequences were longer, were nonexponentially distributed, and resulted in an altered velocity upon restart of translocation downstream of Chi. We propose a model for the recognition of Chi sequences to explain the origin of pausing during failed and successful hotspot recognition.
journal_name
Proc Natl Acad Sci U S Aauthors
Carrasco C,Gilhooly NS,Dillingham MS,Moreno-Herrero Fdoi
10.1073/pnas.1303035110subject
Has Abstractpub_date
2013-07-09 00:00:00pages
E2562-71issue
28eissn
0027-8424issn
1091-6490pii
1303035110journal_volume
110pub_type
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