Direct observation of helicase-topoisomerase coupling within reverse gyrase.
DNA topoisomerase
helicase
magnetic tweezers
single molecule
Journal
Proceedings of the National Academy of Sciences of the United States of America
ISSN: 1091-6490
Titre abrégé: Proc Natl Acad Sci U S A
Pays: United States
ID NLM: 7505876
Informations de publication
Date de publication:
19 05 2020
19 05 2020
Historique:
pubmed:
7
5
2020
medline:
18
8
2020
entrez:
7
5
2020
Statut:
ppublish
Résumé
Reverse gyrases (RGs) are the only topoisomerases capable of generating positive supercoils in DNA. Members of the type IA family, they do so by generating a single-strand break in substrate DNA and then manipulating the two single strands to generate positive topology. Here, we use single-molecule experimentation to reveal the obligatory succession of steps that make up the catalytic cycle of RG. In the initial state, RG binds to DNA and unwinds ∼2 turns of the double helix in an ATP-independent fashion. Upon nucleotide binding, RG then rewinds ∼1 turn of DNA. Nucleotide hydrolysis and/or product release leads to an increase of 2 units of DNA writhe and resetting of the enzyme, for a net change of topology of +1 turn per cycle. Final dissociation of RG from DNA results in rewinding of the 2 turns of DNA that were initially disrupted. These results show how tight coupling of the helicase and topoisomerase activities allows for induction of positive supercoiling despite opposing torque.
Identifiants
pubmed: 32371489
pii: 1921848117
doi: 10.1073/pnas.1921848117
pmc: PMC7245102
doi:
Substances chimiques
DNA, Bacterial
0
DNA-Binding Proteins
0
Adenosine Triphosphate
8L70Q75FXE
DNA
9007-49-2
DNA Helicases
EC 3.6.4.-
DNA Topoisomerases
EC 5.99.1.-
DNA reverse gyrase
EC 5.99.1.-
DNA Topoisomerases, Type I
EC 5.99.1.2
Types de publication
Journal Article
Research Support, Non-U.S. Gov't
Langues
eng
Sous-ensembles de citation
IM
Pagination
10856-10864Informations de copyright
Copyright © 2020 the Author(s). Published by PNAS.
Déclaration de conflit d'intérêts
The authors declare no competing interest.
Références
Nature. 2000 Apr 20;404(6780):901-4
pubmed: 10786800
J Mol Biol. 2013 Jan 9;425(1):32-40
pubmed: 23123378
Phys Chem Chem Phys. 2011 Jun 7;13(21):10009-19
pubmed: 21350762
J Mol Biol. 2011 May 20;408(5):839-49
pubmed: 21435345
Methods Mol Biol. 2018;1703:1-20
pubmed: 29177730
PLoS One. 2014 Jan 02;9(1):e83582
pubmed: 24392087
Cold Spring Harb Protoc. 2012 Jan 01;2012(1):34-49
pubmed: 22194259
Biochimie. 2007 Apr;89(4):447-55
pubmed: 17316953
Nat Genet. 1999 May;22(1):82-4
pubmed: 10319867
Proc Natl Acad Sci U S A. 2002 Sep 17;99(19):12126-31
pubmed: 12167668
Nucleic Acids Res. 2004 Jul 06;32(12):3537-45
pubmed: 15247343
Cell. 2007 Jan 12;128(1):101-14
pubmed: 17218258
Nucleic Acids Res. 2013 Jan;41(2):1058-70
pubmed: 23209025
Nature. 1984 Jun 21-27;309(5970):677-81
pubmed: 6328327
Nucleic Acids Res. 2008 Aug;36(14):4587-97
pubmed: 18614606
EMBO J. 1989 Oct;8(10):3135-9
pubmed: 2555155
J Mol Biol. 1985 Oct 5;185(3):625-37
pubmed: 2997454
Nucleic Acids Res. 2014 Jul;42(12):7935-46
pubmed: 24880688
Nat Struct Mol Biol. 2019 Apr;26(4):267-274
pubmed: 30936532
Mol Cell. 2012 Sep 28;47(6):886-96
pubmed: 22885009
Proc Natl Acad Sci U S A. 2015 Jun 16;112(24):7495-500
pubmed: 26023188
Nature. 2006 Jan 5;439(7072):100-104
pubmed: 16397501
Cell. 1995 Nov 17;83(4):655-66
pubmed: 7585968
Nucleic Acids Res. 2011 Mar;39(5):1789-800
pubmed: 21051354
Annu Rev Biophys Biomol Struct. 2005;34:201-19
pubmed: 15869388
Biochimie. 2007 Apr;89(4):456-67
pubmed: 17141394
J Bacteriol. 2004 Jul;186(14):4829-33
pubmed: 15231817
FEBS J. 2016 Apr;283(8):1372-84
pubmed: 26836040
Nat Struct Mol Biol. 2015 Jun;22(6):452-7
pubmed: 25961799
Mol Microbiol. 2020 Feb;113(2):356-368
pubmed: 31713907
Nat Struct Mol Biol. 2017 May;24(5):484-490
pubmed: 28414321
J Biol Chem. 2006 Mar 3;281(9):5640-7
pubmed: 16407212
Nature. 2007 Jul 12;448(7150):213-7
pubmed: 17589503
Science. 2006 Nov 17;314(5802):1139-43
pubmed: 17110577
Proc Natl Acad Sci U S A. 1998 Dec 22;95(26):15218-22
pubmed: 9860949
Proc Natl Acad Sci U S A. 1978 Aug;75(8):3557-61
pubmed: 16592550
Science. 2011 Mar 11;331(6022):1328-32
pubmed: 21393545
Extremophiles. 2008 Nov;12(6):799-809
pubmed: 18777006
Science. 1996 Mar 29;271(5257):1835-7
pubmed: 8596951
Nat Struct Mol Biol. 2018 Dec;25(12):1111-1118
pubmed: 30478267
Nat Chem Biol. 2018 Jun;14(6):565-574
pubmed: 29662209
EMBO J. 1985 Oct;4(10):2705-10
pubmed: 16453636
J Mol Biol. 2003 May 30;329(2):271-82
pubmed: 12758075
Proc Natl Acad Sci U S A. 2004 Apr 6;101(14):4776-80
pubmed: 15037753
J Biol Chem. 2000 Jun 30;275(26):19498-504
pubmed: 10748189
J Mol Biol. 1968 Apr 14;33(1):141-71
pubmed: 4296517
Eur J Biochem. 1999 Feb;260(1):103-11
pubmed: 10091589
Biophys J. 2005 Jun;88(6):4124-36
pubmed: 15778439
Proc Natl Acad Sci U S A. 2013 Sep 17;110(38):E3587-94
pubmed: 24003117
J Biol Chem. 2003 Oct 24;278(43):42668-78
pubmed: 12909639
EMBO J. 1985 Aug;4(8):2123-8
pubmed: 14708549
Biochemistry. 1988 Dec 27;27(26):9102-8
pubmed: 2853975
J Biol Chem. 2000 Mar 31;275(13):9636-44
pubmed: 10734115
Science. 1996 Apr 12;272(5259):258-62
pubmed: 8602509
Biophys J. 2005 Jul;89(1):384-92
pubmed: 15863484
Nat Struct Mol Biol. 2014 Mar;21(3):261-8
pubmed: 24509834
Nature. 2012 Oct 18;490(7420):431-4
pubmed: 22960746
J Mol Biol. 1987 May 5;195(1):205-13
pubmed: 3656410
Mol Cell. 2008 Jun 20;30(6):779-89
pubmed: 18570879
Nature. 2016 Aug 11;536(7615):234-7
pubmed: 27487215
Nat Struct Mol Biol. 2012 Apr 08;19(5):538-46, S1
pubmed: 22484318
Nucleic Acids Res. 2009 Jul;37(13):4287-95
pubmed: 19443439