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ssDNA reeling is an intermediate step in the reiterative DNA unwinding activity of the WRN-1 helicase

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Title
ssDNA reeling is an intermediate step in the reiterative DNA unwinding activity of the WRN-1 helicase
Author(s)
Le, Son Truong; Choi, Seoyun; Lee, Seung-Won; Hajin Kim; Ahn, Byungchan
Publication Date
2023-09
Journal
Journal of Biological Chemistry, v.299, no.9
Publisher
American Society for Biochemistry and Molecular Biology Inc.
Abstract
RecQ helicases are highly conserved between bacteria and humans. These helicases unwind various DNA structures in the 3′ to 5′. Defective helicase activity elevates genomic instability and is associated with predisposition to cancer and/or premature aging. Recent single-molecule analyses have revealed the repetitive unwinding behavior of RecQ helicases from Escherichia coli to humans. However, the detailed mechanisms underlying this behavior are unclear. Here, we performed single-molecule studies of WRN-1 Caenorhabditis elegans RecQ helicase on various DNA constructs and characterized WRN-1 unwinding dynamics. We showed that WRN-1 persistently repeated cycles of DNA unwinding and rewinding with an unwinding limit of 25 to 31 bp per cycle. Furthermore, by monitoring the ends of the displaced strand during DNA unwinding we demonstrated that WRN-1 reels in the ssDNA overhang in an ATP-dependent manner. While WRN-1 reeling activity was inhibited by a C. elegans homolog of human replication protein A, we found that C. elegans replication protein A actually switched the reiterative unwinding activity of WRN-1 to unidirectional unwinding. These results reveal that reeling-in ssDNA is an intermediate step in the reiterative unwinding process for WRN-1 (i.e., the process proceeds via unwinding-reeling-rewinding). We propose that the reiterative unwinding activity of WRN-1 may prevent extensive unwinding, allow time for partner proteins to assemble on the active region, and permit additional modulation in vivo.
URI
https://pr.ibs.re.kr/handle/8788114/14407
DOI
10.1016/j.jbc.2023.105081
ISSN
0021-9258
Appears in Collections:
Center for Genomic Integrity(유전체 항상성 연구단) > 1. Journal Papers (저널논문)
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