Deoxyuridine in DNA has an inhibitory and promutagenic effect on RNA transcription by diverse RNA polymerases.
Base Pairing
Base Sequence
Cell Line, Tumor
DNA
/ genetics
DNA Repair
DNA Replication
DNA-Directed RNA Polymerases
/ genetics
Deoxyuridine
/ metabolism
Fibroblasts
/ cytology
Humans
Kinetics
Mutation
Promoter Regions, Genetic
Protein Binding
RNA
/ genetics
RNA Polymerase II
/ genetics
Thermodynamics
Transcription, Genetic
Viral Proteins
/ genetics
Journal
Nucleic acids research
ISSN: 1362-4962
Titre abrégé: Nucleic Acids Res
Pays: England
ID NLM: 0411011
Informations de publication
Date de publication:
07 05 2019
07 05 2019
Historique:
accepted:
09
03
2019
revised:
04
03
2019
received:
01
02
2019
pubmed:
21
3
2019
medline:
26
11
2019
entrez:
21
3
2019
Statut:
ppublish
Résumé
dUTP is a close structural congener of dTTP and can be readily incorporated into DNA opposite to adenine during DNA replication leading to non-mutagenic dU/A base pairs ('uracilation'). We find that dU/A pairs located within DNA transcriptional templates optimized for either T7 RNA polymerase (T7 RNAP) or human RNA polymerase II (pol II) have inhibitory and mutagenic effects on transcription. The data for T7 RNAP establishes that even a single dU/A pair can inhibit promoter binding and transcription initiation up to 30-fold, and that inhibitory effects on transcription elongation are also possible. Sequencing of the mRNA transcribed from uniformly uracilated DNA templates by T7 RNAP indicated an increased frequency of transversion and insertion mutations compared to all T/A templates. Strong effects of dU/A pairs on cellular transcription activity and fidelity were also observed with RNA pol II using uracil base excision repair (UBER)-deficient human cells. At the highest levels of template uracilation, transcription by RNA pol II was completely blocked. We propose that these effects arise from the decreased thermodynamic stability and increased dynamics of dU/A pairs in DNA. The potential implications of these findings on gene regulation and disease are discussed.
Identifiants
pubmed: 30892639
pii: 5403496
doi: 10.1093/nar/gkz183
pmc: PMC6486633
doi:
Substances chimiques
Viral Proteins
0
RNA
63231-63-0
DNA
9007-49-2
RNA Polymerase II
EC 2.7.7.-
bacteriophage T7 RNA polymerase
EC 2.7.7.-
DNA-Directed RNA Polymerases
EC 2.7.7.6
Deoxyuridine
W78I7AY22C
Types de publication
Journal Article
Research Support, N.I.H., Extramural
Research Support, Non-U.S. Gov't
Langues
eng
Sous-ensembles de citation
IM
Pagination
4153-4168Subventions
Organisme : NIAID NIH HHS
ID : F32 AI150561
Pays : United States
Organisme : NIAID NIH HHS
ID : R01 AI124777
Pays : United States
Organisme : NCI NIH HHS
ID : T32 CA009110
Pays : United States
Informations de copyright
© The Author(s) 2019. Published by Oxford University Press on behalf of Nucleic Acids Research.
Références
Proc Natl Acad Sci U S A. 2016 Mar 22;113(12):3311-6
pubmed: 26884158
J Biol Chem. 2002 Aug 9;277(32):28491-7
pubmed: 12029089
J Biol Chem. 2015 Feb 27;290(9):5502-11
pubmed: 25572391
Retrovirology. 2008 Jun 05;5:45
pubmed: 18533995
Mol Cell. 2009 Jun 26;34(6):710-21
pubmed: 19560423
Mutat Res. 2000 Aug 30;460(3-4):165-81
pubmed: 10946227
Biochemistry. 2002 Jun 25;41(25):8093-102
pubmed: 12069602
Nucleic Acids Res. 2017 Dec 1;45(21):12413-12424
pubmed: 29036472
Biochemistry. 1993 Apr 27;32(16):4270-4
pubmed: 8476855
EMBO J. 2001 Nov 15;20(22):6520-9
pubmed: 11707422
Cell. 2013 Oct 24;155(3):540-51
pubmed: 24243014
DNA Repair (Amst). 2018 Oct;70:18-24
pubmed: 30098578
Elife. 2016 Sep 20;5:
pubmed: 27644592
J Biol Chem. 2003 Mar 7;278(10):7902-9
pubmed: 12458216
Proc Natl Acad Sci U S A. 2017 Mar 7;114(10):2604-2609
pubmed: 28143930
Curr Opin Genet Dev. 2017 Feb;42:1-7
pubmed: 27939598
J Mol Recognit. 1991 Jul-Dec;4(4):133-41
pubmed: 1665978
J Biol Chem. 2002 Jan 25;277(4):2987-91
pubmed: 11704669
DNA Repair (Amst). 2014 Jul;19:38-47
pubmed: 24746924
J Biol Chem. 2011 Jul 15;286(28):25047-55
pubmed: 21454906
Biochemistry. 2011 Feb 8;50(5):612-7
pubmed: 21190322
Mol Biol Int. 2014;2014:287430
pubmed: 25197572
Science. 1998 Jan 9;279(5348):242-7
pubmed: 9422699
Nature. 2013 Oct 24;502(7472):472-9
pubmed: 24153300
Proc Natl Acad Sci U S A. 2011 May 31;108(22):9244-9
pubmed: 21576478
Nat Chem Biol. 2018 Jul;14(7):680-687
pubmed: 29785056
Biochemistry. 1986 Oct 7;25(20):5920-8
pubmed: 3098280
J Virol Methods. 2008 Aug;151(2):217-24
pubmed: 18571738
Science. 2011 Jul 1;333(6038):53-8
pubmed: 21596952
Curr Opin Microbiol. 2013 Apr;16(2):163-70
pubmed: 23541339
PLoS Genet. 2012;8(6):e1002738
pubmed: 22685418
Biomol Detect Quantif. 2014 Dec 04;2:11-29
pubmed: 27896140
Biochemistry. 1993 Apr 27;32(16):4281-5
pubmed: 8476857
Mol Gen Genet. 1981;183(3):561-3
pubmed: 7038382
J Am Chem Soc. 2012 May 16;134(19):8231-40
pubmed: 22509745
Nucleic Acids Res. 2013 Oct;41(19):9090-104
pubmed: 23925128
Biochemistry. 2018 Oct 2;57(39):5666-5671
pubmed: 30185020
Mol Biol Evol. 1987 May;4(3):203-21
pubmed: 3328815
Proc Natl Acad Sci U S A. 2013 Feb 5;110(6):E448-57
pubmed: 23341616
Elife. 2015 Dec 10;4:
pubmed: 26652005
Mutat Res. 2002 Dec 29;510(1-2):131-40
pubmed: 12459449
Nature. 1999 May 6;399(6731):80-3
pubmed: 10331394
Biochemistry. 1994 Jun 7;33(22):6918-24
pubmed: 7911327
Mol Cell Biol. 1989 Feb;9(2):469-76
pubmed: 2469002
J Biol Chem. 2003 Feb 28;278(9):7294-9
pubmed: 12466278
Nucleic Acids Res. 2015 Jan;43(Database issue):D298-9
pubmed: 25378308
J Biol Chem. 2017 Nov 3;292(44):18145-18160
pubmed: 28882896