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Title: Mechanism of error-free DNA synthesis across N1-methyl-deoxyadenosine by human DNA polymerase-ι

Abstract

N1-methyl-deoxyadenosine (1-MeA) is formed by methylation of deoxyadenosine at the N1 atom. 1-MeA presents a block to replicative DNA polymerases due to its inability to participate in Watson-Crick (W-C) base pairing. Here we determine how human DNA polymerase-ι (Polι) promotes error-free replication across 1-MeA. Steady state kinetic analyses indicate that Polι is ~100 fold more efficient in incorporating the correct nucleotide T versus the incorrect nucleotide C opposite 1-MeA. To understand the basis of this selectivity, we determined ternary structures of Polι bound to template 1-MeA and incoming dTTP or dCTP. In both structures, template 1-MeA rotates to the syn conformation but pairs differently with dTTP versus dCTP. Thus, whereas dTTP partakes in stable Hoogsteen base pairing with 1-MeA, dCTP fails to gain a “foothold” and is largely disordered. Together, our kinetic and structural studies show how Polι maintains discrimination between correct and incorrect incoming nucleotide opposite 1-MeA in preserving genome integrity.

Authors:
; ; ; ; ; ;
Publication Date:
Research Org.:
Brookhaven National Lab. (BNL), Upton, NY (United States)
Sponsoring Org.:
USDOE Office of Science (SC), Basic Energy Sciences (BES) (SC-22)
OSTI Identifier:
1409564
Report Number(s):
BNL-114616-2017-JA¿¿¿
Journal ID: ISSN 2045-2322
DOE Contract Number:  
SC0012704
Resource Type:
Journal Article
Journal Name:
Scientific Reports
Additional Journal Information:
Journal Volume: 7; Journal ID: ISSN 2045-2322
Publisher:
Nature Publishing Group
Country of Publication:
United States
Language:
English
Subject:
59 BASIC BIOLOGICAL SCIENCES; 60 APPLIED LIFE SCIENCES

Citation Formats

Jain, Rinku, Choudhury, Jayati Roy, Buku, Angeliki, Johnson, Robert E., Prakash, Louise, Prakash, Satya, and Aggarwal, Aneel K. Mechanism of error-free DNA synthesis across N1-methyl-deoxyadenosine by human DNA polymerase-ι. United States: N. p., 2017. Web. doi:10.1038/srep43904.
Jain, Rinku, Choudhury, Jayati Roy, Buku, Angeliki, Johnson, Robert E., Prakash, Louise, Prakash, Satya, & Aggarwal, Aneel K. Mechanism of error-free DNA synthesis across N1-methyl-deoxyadenosine by human DNA polymerase-ι. United States. doi:10.1038/srep43904.
Jain, Rinku, Choudhury, Jayati Roy, Buku, Angeliki, Johnson, Robert E., Prakash, Louise, Prakash, Satya, and Aggarwal, Aneel K. Wed . "Mechanism of error-free DNA synthesis across N1-methyl-deoxyadenosine by human DNA polymerase-ι". United States. doi:10.1038/srep43904.
@article{osti_1409564,
title = {Mechanism of error-free DNA synthesis across N1-methyl-deoxyadenosine by human DNA polymerase-ι},
author = {Jain, Rinku and Choudhury, Jayati Roy and Buku, Angeliki and Johnson, Robert E. and Prakash, Louise and Prakash, Satya and Aggarwal, Aneel K.},
abstractNote = {N1-methyl-deoxyadenosine (1-MeA) is formed by methylation of deoxyadenosine at the N1 atom. 1-MeA presents a block to replicative DNA polymerases due to its inability to participate in Watson-Crick (W-C) base pairing. Here we determine how human DNA polymerase-ι (Polι) promotes error-free replication across 1-MeA. Steady state kinetic analyses indicate that Polι is ~100 fold more efficient in incorporating the correct nucleotide T versus the incorrect nucleotide C opposite 1-MeA. To understand the basis of this selectivity, we determined ternary structures of Polι bound to template 1-MeA and incoming dTTP or dCTP. In both structures, template 1-MeA rotates to the syn conformation but pairs differently with dTTP versus dCTP. Thus, whereas dTTP partakes in stable Hoogsteen base pairing with 1-MeA, dCTP fails to gain a “foothold” and is largely disordered. Together, our kinetic and structural studies show how Polι maintains discrimination between correct and incorrect incoming nucleotide opposite 1-MeA in preserving genome integrity.},
doi = {10.1038/srep43904},
journal = {Scientific Reports},
issn = {2045-2322},
number = ,
volume = 7,
place = {United States},
year = {2017},
month = {3}
}

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