Structural insights into dynamics of RecU-HJ complex formation elucidates key role of NTR and stalk region toward formation of reactive state.

Abstract:

:Holliday junction (HJ) resolving enzyme RecU is involved in DNA repair and recombination. We have determined the crystal structure of inactive mutant (D88N) of RecU from Bacillus subtilis in complex with a 12 base palindromic DNA fragment at a resolution of 3.2 Å. This structure shows the stalk region and the essential N-terminal region (NTR) previously unseen in our DNA unbound structure. The flexible nature of the NTR in solution was confirmed using SAXS. Thermofluor studies performed to assess the stability of RecU in complex with the arms of an HJ indicate that it confers stability. Further, we performed molecular dynamics (MD) simulations of wild type and an NTR deletion variant of RecU, with and without HJ. The NTR is observed to be highly flexible in simulations of the unbound RecU, in agreement with SAXS observations. These simulations revealed domain dynamics of RecU and their role in the formation of complex with HJ. The MD simulations also elucidate key roles of the NTR, stalk region, and breathing motion of RecU in the formation of the reactive state.

journal_name

Nucleic Acids Res

journal_title

Nucleic acids research

authors

Khavnekar S,Dantu SC,Sedelnikova S,Ayora S,Rafferty J,Kale A

doi

10.1093/nar/gkw1165

subject

Has Abstract

pub_date

2017-01-25 00:00:00

pages

975-986

issue

2

eissn

0305-1048

issn

1362-4962

pii

gkw1165

journal_volume

45

pub_type

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