Molecular insights into replication initiation in a multipartite genome harboring bacterium Deinococcus radiodurans.
ATPase
DNA–protein interaction
bacterial genetics
chromosomes
metabolic regulation
origin of replication
protein–protein interaction
radiation biology
Journal
The Journal of biological chemistry
ISSN: 1083-351X
Titre abrégé: J Biol Chem
Pays: United States
ID NLM: 2985121R
Informations de publication
Date de publication:
Historique:
received:
19
11
2020
revised:
12
02
2021
accepted:
18
02
2021
pubmed:
25
2
2021
medline:
31
8
2021
entrez:
24
2
2021
Statut:
ppublish
Résumé
Deinococcus radiodurans harbors a multipartite ploid genome system consisting of two chromosomes and two plasmids present in multiple copies. How these discrete genome elements are maintained and inherited is not well understood. PprA, a pleiotropic protein involved in radioresistance, has been characterized for its roles in DNA repair, genome segregation, and cell division in this bacterium. Here, we show that PprA regulates ploidy of chromosome I and II and inhibits the activity of drDnaA, the initiator protein in D. radiodurans. We found that pprA deletion resulted in an increased genomic content and ploidy of both the chromosomal elements. Expression of PprA in trans rescued the phenotypes of the pprA mutant. To understand the molecular mechanism underlying these phenotypes, we characterized drDnaA and drDnaB. As expected for an initiator protein, recombinant drDnaA showed sequence-specific interactions with the putative oriC sequence in chromosome I (oriCI). Both drDnaA and drDnaB showed ATPase activity, also typical of initiator proteins, but only drDnaB exhibited 5'→3' dsDNA helicase activity in vitro. drDnaA and drDnaB showed homotypic and heterotypic interactions with each other, which were perturbed by PprA. Interestingly, PprA has inhibited the ATPase activity of drDnaA but showed no effect on the activity of drDnaB. Regulation of chromosome copy number and inhibition of the initiator protein functions by PprA strongly suggest that it plays a role as a checkpoint regulator of the DNA replication initiation in D. radiodurans perhaps through its interaction with the replication initiation machinery.
Identifiants
pubmed: 33626388
pii: S0021-9258(21)00224-6
doi: 10.1016/j.jbc.2021.100451
pmc: PMC7988490
pii:
doi:
Substances chimiques
Bacterial Proteins
0
DNA Gyrase
EC 5.99.1.3
Types de publication
Journal Article
Langues
eng
Sous-ensembles de citation
IM
Pagination
100451Informations de copyright
Copyright © 2021. Published by Elsevier Inc.
Déclaration de conflit d'intérêts
Conflict of interest Authors have no financial or nonfinancial competing interests.
Références
EMBO J. 1995 May 1;14(9):2106-11
pubmed: 7744016
J Bacteriol. 2003 Nov;185(22):6633-9
pubmed: 14594837
Proc Natl Acad Sci U S A. 1998 Jan 6;95(1):120-5
pubmed: 9419339
Front Microbiol. 2015 Jun 02;6:545
pubmed: 26082765
Appl Microbiol Biotechnol. 2006 Oct;72(4):790-6
pubmed: 16586106
EMBO J. 2001 Jun 15;20(12):3262-71
pubmed: 11406602
FASEB J. 2019 Mar;33(3):3647-3658
pubmed: 30481062
Cell. 2008 Oct 3;135(1):74-84
pubmed: 18854156
Protein Sci. 2014 Oct;23(10):1349-58
pubmed: 25044036
Mol Microbiol. 2005 Jun;56(5):1129-38
pubmed: 15882408
J Bacteriol. 2009 Sep;191(17):5458-70
pubmed: 19542275
PLoS One. 2014 Jan 15;9(1):e85288
pubmed: 24454836
Clin Chem. 2009 Apr;55(4):611-22
pubmed: 19246619
Nucleic Acids Res. 2002 Sep 15;30(18):4051-60
pubmed: 12235389
Biochim Biophys Acta. 2012 Jul;1819(7):826-9
pubmed: 22306663
Mol Microbiol. 2006 Feb;59(4):1308-16
pubmed: 16430702
Mol Microbiol. 2004 Oct;54(1):278-85
pubmed: 15458422
Curr Opin Microbiol. 2014 Dec;22:120-6
pubmed: 25460805
FEMS Microbiol Rev. 2002 Nov;26(4):355-74
pubmed: 12413665
Nucleic Acids Res. 2003 Apr 15;31(8):2077-86
pubmed: 12682358
EMBO J. 2007 Jul 11;26(13):3124-31
pubmed: 17557077
J Genet. 2018 Sep;97(4):1013-1038
pubmed: 30262715
Biochim Biophys Acta. 2012 Jul;1820(7):1052-61
pubmed: 22503789
Nucleic Acids Res. 2001 Jun 1;29(11):2251-9
pubmed: 11376143
Front Microbiol. 2015 Dec 10;6:1370
pubmed: 26696980
Mol Microbiol. 2011 Feb;79(4):1089-100
pubmed: 21235642
Mol Microbiol. 2000 May;36(3):557-69
pubmed: 10844646
J Bacteriol. 2003 Nov;185(21):6316-24
pubmed: 14563866
Proc Natl Acad Sci U S A. 1998 May 12;95(10):5752-6
pubmed: 9576956
Int J Biochem Cell Biol. 2013 Nov;45(11):2541-52
pubmed: 23994692
Microbiol Res. 2019 Jun - Aug;223-225:22-32
pubmed: 31178048
FEMS Microbiol Rev. 2007 Jul;31(4):378-87
pubmed: 17459114
J Bacteriol. 1992 May;174(10):3220-6
pubmed: 1577691
J Bacteriol. 2010 Aug;192(15):3903-14
pubmed: 20511501
Nucleic Acids Res. 2003 Dec 1;31(23):6828-40
pubmed: 14627816
Curr Opin Chem Biol. 2011 Oct;15(5):587-94
pubmed: 21855395
Methods. 2012 Dec;58(4):325-34
pubmed: 22841567
Photochem Photobiol. 2000 May;71(5):514-23
pubmed: 10818781
Nat Rev Microbiol. 2007 May;5(5):343-54
pubmed: 17435790
J Mol Biol. 2000 Jun 9;299(3):655-65
pubmed: 10835275
PLoS One. 2006 Dec 20;1:e92
pubmed: 17183724
Cell. 2003 Aug 22;114(4):521-30
pubmed: 12941279
Curr Microbiol. 1994 Jul;29(1):23-9
pubmed: 7764984
mSphere. 2016 Jan 13;1(1):
pubmed: 27303692
Biochem J. 2019 Mar 15;476(5):909-930
pubmed: 30804051
Eur J Biochem. 1999 Mar;260(2):325-35
pubmed: 10095766
mSphere. 2018 Jul 18;3(4):
pubmed: 30021877
Science. 1999 Nov 19;286(5444):1571-7
pubmed: 10567266
Microbiol Mol Biol Rev. 2016 Nov 30;81(1):
pubmed: 27903655
Mol Microbiol. 2019 Jan;111(1):118-130
pubmed: 30285297
Genome Res. 2004 Jun;14(6):1188-90
pubmed: 15173120
FEBS J. 2012 Jan;279(2):234-50
pubmed: 22074440
Annu Rev Biochem. 2000;69:651-97
pubmed: 10966472
J Mol Biol. 1997 Dec 12;274(4):546-61
pubmed: 9417934
Biochimie. 1999 Aug-Sep;81(8-9):819-25
pubmed: 10572294
Life Sci Alliance. 2020 Nov 16;4(1):
pubmed: 33199509
PLoS One. 2014 Dec 31;9(12):e115918
pubmed: 25551229
Extremophiles. 2016 Mar;20(2):195-205
pubmed: 26847200
Nature. 2006 Oct 5;443(7111):569-73
pubmed: 17006450
Cell. 2010 Jun 11;141(6):1088, 1088.e1
pubmed: 20550941
Mol Microbiol. 2019 Sep;112(3):854-865
pubmed: 31162841
Front Microbiol. 2018 Nov 30;9:2932
pubmed: 30559732
Microbiol Mol Biol Rev. 2011 Mar;75(1):133-91
pubmed: 21372322
Mol Microbiol. 2000 Sep;37(5):1270-9
pubmed: 10972842
Nucleic Acids Res. 2019 Jan 8;47(D1):D74-D77
pubmed: 30364951
DNA Repair (Amst). 2013 Apr 1;12(4):265-72
pubmed: 23403184
FEBS J. 2014 Feb;281(4):1256-66
pubmed: 24387047
Front Microbiol. 2018 Aug 07;9:1833
pubmed: 30131796
J Biol Chem. 1986 Apr 5;261(10):4738-48
pubmed: 3007474
EMBO J. 1988 Sep;7(9):2911-7
pubmed: 2846289
J Bacteriol. 2009 Jan;191(1):249-60
pubmed: 18931108
Genes (Basel). 2017 May 10;8(5):
pubmed: 28489024
Biochem J. 2004 Aug 15;382(Pt 1):247-52
pubmed: 15137907
Microbiology (Reading). 2016 Aug;162(8):1321-1334
pubmed: 27368754
Nat Rev Microbiol. 2005 Nov;3(11):882-92
pubmed: 16261171
Sci Adv. 2016 Apr 22;2(4):e1501914
pubmed: 27152358
J Bacteriol. 1978 Apr;134(1):71-5
pubmed: 649572
Cold Spring Harb Perspect Biol. 2013 Apr 01;5(4):a012922
pubmed: 23471435