Single-molecule Studies Reveal Reciprocating of WRN Helicase Core Along SsDNA During DNA Unwinding

Overview

Journal Sci Rep

Specialty Science

Date 2017 Mar 8

PMID 28266653

Citations 22

Authors

Wen-Qiang Wu

Xi-Miao Hou

Bo Zhang

Philippe Fosse

Brigitte Rene

Olivier Mauffret

Ming Li

Shuo-Xing Dou

Xu-Guang Xi

Affiliations

Soon will be listed here.

Abstract

Werner syndrome is caused by mutations in the WRN gene encoding WRN helicase. A knowledge of WRN helicase's DNA unwinding mechanism in vitro is helpful for predicting its behaviors in vivo, and then understanding their biological functions. In the present study, for deeply understanding the DNA unwinding mechanism of WRN, we comprehensively characterized the DNA unwinding properties of chicken WRN helicase core in details, by taking advantages of single-molecule fluorescence resonance energy transfer (smFRET) method. We showed that WRN exhibits repetitive DNA unwinding and translocation behaviors on different DNA structures, including forked, overhanging and G-quadruplex-containing DNAs with an apparently limited unwinding processivity. It was further revealed that the repetitive behaviors were caused by reciprocating of WRN along the same ssDNA, rather than by complete dissociation from and rebinding to substrates or by strand switching. The present study sheds new light on the mechanism for WRN functioning.

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References

Dallaire A, Garand C, Paquel E, Mitchell S, de Cabo R, Simard M . Down regulation of miR-124 in both Werner syndrome DNA helicase mutant mice and mutant Caenorhabditis elegans wrn-1 reveals the importance of this microRNA in accelerated aging. Aging (Albany NY). 2012; 4(9):636-47. PMC: 3492227. DOI: 10.18632/aging.100489. View

Gray M, Shen J, Blank A, Sopher B, Martin G, Oshima J . The Werner syndrome protein is a DNA helicase. Nat Genet. 1997; 17(1):100-3. DOI: 10.1038/ng0997-100. View

Yang Y, Dou S, Xu Y, Bazeille N, Wang P, Rigolet P . Kinetic mechanism of DNA unwinding by the BLM helicase core and molecular basis for its low processivity. Biochemistry. 2009; 49(4):656-68. DOI: 10.1021/bi901459c. View

Klaue D, Kobbe D, Kemmerich F, Kozikowska A, Puchta H, Seidel R . Fork sensing and strand switching control antagonistic activities of RecQ helicases. Nat Commun. 2013; 4:2024. PMC: 3709500. DOI: 10.1038/ncomms3024. View

Shen J, Loeb L, Fry M . Werner syndrome protein. II. Characterization of the integral 3' --> 5' DNA exonuclease. J Biol Chem. 1998; 273(51):34145-50. DOI: 10.1074/jbc.273.51.34145. View

Brosh Jr R, Bohr V . Human premature aging, DNA repair and RecQ helicases. Nucleic Acids Res. 2007; 35(22):7527-44. PMC: 2190726. DOI: 10.1093/nar/gkm1008. View

Imamura O, Fujita K, Itoh C, Takeda S, Furuichi Y, Matsumoto T . Werner and Bloom helicases are involved in DNA repair in a complementary fashion. Oncogene. 2002; 21(6):954-63. DOI: 10.1038/sj.onc.1205143. View

Roy R, Kozlov A, Lohman T, Ha T . SSB protein diffusion on single-stranded DNA stimulates RecA filament formation. Nature. 2009; 461(7267):1092-7. PMC: 2782680. DOI: 10.1038/nature08442. View

Tippana R, Hwang H, Opresko P, Bohr V, Myong S . Single-molecule imaging reveals a common mechanism shared by G-quadruplex-resolving helicases. Proc Natl Acad Sci U S A. 2016; 113(30):8448-53. PMC: 4968719. DOI: 10.1073/pnas.1603724113. View

10.

Menetski J, Kowalczykowski S . Interaction of recA protein with single-stranded DNA. Quantitative aspects of binding affinity modulation by nucleotide cofactors. J Mol Biol. 1985; 181(2):281-95. DOI: 10.1016/0022-2836(85)90092-0. View

11.

Bernstein D, Zittel M, Keck J . High-resolution structure of the E.coli RecQ helicase catalytic core. EMBO J. 2003; 22(19):4910-21. PMC: 204483. DOI: 10.1093/emboj/cdg500. View

12.

Wu W, Hou X, Li M, Dou S, Xi X . BLM unfolds G-quadruplexes in different structural environments through different mechanisms. Nucleic Acids Res. 2015; 43(9):4614-26. PMC: 4482088. DOI: 10.1093/nar/gkv361. View

13.

Rasnik I, Myong S, Cheng W, Lohman T, Ha T . DNA-binding orientation and domain conformation of the E. coli rep helicase monomer bound to a partial duplex junction: single-molecule studies of fluorescently labeled enzymes. J Mol Biol. 2004; 336(2):395-408. DOI: 10.1016/j.jmb.2003.12.031. View

14.

Mendoza O, Bourdoncle A, Boule J, Brosh Jr R, Mergny J . G-quadruplexes and helicases. Nucleic Acids Res. 2016; 44(5):1989-2006. PMC: 4797304. DOI: 10.1093/nar/gkw079. View

15.

Zhang B, Wu W, Liu N, Duan X, Li M, Dou S . G-quadruplex and G-rich sequence stimulate Pif1p-catalyzed downstream duplex DNA unwinding through reducing waiting time at ss/dsDNA junction. Nucleic Acids Res. 2016; 44(17):8385-94. PMC: 5041479. DOI: 10.1093/nar/gkw669. View

16.

Wang S, Qin W, Li J, Lu Y, Lu K, Nong D . Unwinding forward and sliding back: an intermittent unwinding mode of the BLM helicase. Nucleic Acids Res. 2015; 43(7):3736-46. PMC: 4402530. DOI: 10.1093/nar/gkv209. View

17.

Lee S, Syed S, Enemark E, Schuck S, Stenlund A, Ha T . Dynamic look at DNA unwinding by a replicative helicase. Proc Natl Acad Sci U S A. 2014; 111(9):E827-35. PMC: 3948270. DOI: 10.1073/pnas.1322254111. View

18.

Kobbe D, Blanck S, Demand K, Focke M, Puchta H . AtRECQ2, a RecQ helicase homologue from Arabidopsis thaliana, is able to disrupt various recombinogenic DNA structures in vitro. Plant J. 2008; 55(3):397-405. DOI: 10.1111/j.0960-7412.2008.03511.x. View

19.

Park J, Myong S, Niedziela-Majka A, Lee K, Yu J, Lohman T . PcrA helicase dismantles RecA filaments by reeling in DNA in uniform steps. Cell. 2010; 142(4):544-55. PMC: 2943210. DOI: 10.1016/j.cell.2010.07.016. View

20.

Mohaghegh P, Karow J, Brosh Jr R, Bohr V, Hickson I . The Bloom's and Werner's syndrome proteins are DNA structure-specific helicases. Nucleic Acids Res. 2001; 29(13):2843-9. PMC: 55766. DOI: 10.1093/nar/29.13.2843. View