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B Family DNA Polymerases Asymmetrically Recognize Pyrimidines and Purines
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SYSNO ASEP 0364221 Document Type J - Journal Article R&D Document Type Journal Article Subsidiary J Článek ve WOS Title B Family DNA Polymerases Asymmetrically Recognize Pyrimidines and Purines Author(s) Lund, T. J. (US)
Cavanaugh, N. A. (US)
Joubert, Nicolas (UOCHB-X)
Urban, M. (US)
Patro, J. N. (US)
Hocek, Michal (UOCHB-X) RID, ORCID
Kuchta, R. D. (US)Number of authors 7 Source Title Biochemistry. - : American Chemical Society - ISSN 0006-2960
Roč. 50, č. 33 (2011), s. 7243-7250Number of pages 8 s. Language eng - English Country US - United States Keywords active-site tightness ; genetic alphabet ; deoxynucleoside ; phosphoramidites Subject RIV CC - Organic Chemistry R&D Projects LC512 GA MŠMT - Ministry of Education, Youth and Sports (MEYS) IAA400550902 GA AV ČR - Academy of Sciences of the Czech Republic (AV ČR) CEZ AV0Z40550506 - UOCHB-X (2005-2011) UT WOS 000294076100018 DOI 10.1021/bi2006916 Annotation We utilized a series of modified pyrimidine analogues to determine the mechanism used by human DNA polymerase α and herpes simplex virus I DNA polymerase to polymerize pyrimidine dNTPs. Removing O2 of a pyrimidine dNTP vastly decreased incorporation by these enzymes and also compromised fidelity in the case of C analogues, while removing O2 from the templating base had more modest effects. Removing the Watson-Crick hydrogen bonding groups greatly impaired polymerization. The Watson-Crick hydrogen bonding plays an important role in enhancing correct dNTP polymerization, but are not essential for preventing misincorporation. These studies also indicate that DNA polymerases recognize bases extremely asymmetrically, both in terms of whether they are a purine or pyrimidine and whether they are in the template or are the incoming dNTP. The mechanistic implications of these results regarding how polymerases discriminate between right and wrong dNTPs are discussed. Workplace Institute of Organic Chemistry and Biochemistry Contact asep@uochb.cas.cz ; Kateřina Šperková, Tel.: 232 002 584 ; Viktorie Chládková, Tel.: 232 002 434 Year of Publishing 2012
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