Journal
JOURNAL OF MOLECULAR BIOLOGY
Volume 395, Issue 2, Pages 282-290Publisher
ACADEMIC PRESS LTD- ELSEVIER SCIENCE LTD
DOI: 10.1016/j.jmb.2009.11.003
Keywords
DNA polymerase lambda; sugar selection; pre-steady-state kinetics; nucleotide analogs; steric gate
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Funding
- National Institutes of Health [GM079403, T32-GM008512-13]
- American Heart Association [0815382D, 0415129B, 0615091B]
- International P.E.O. Scholar Award
- Ohio State University
- National Science Foundation [MCB-0447899]
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During DNA synthesis, most DNA polymerases and reverse transcriptases select against ribonucleotides via a steric clash between the ribose 2'-hydroxyl group and the bulky side chain of an active-site residue. In this study, we demonstrated that human DNA polymerase lambda used a novel sugar selection mechanism to discriminate against ribonucleotides, whereby the ribose 2'-hydroxyl group was excluded mostly by a backbone segment and slightly by the side chain of Y505. Such steric clash was further demonstrated to be dependent on the size and orientation of the substituent covalently attached at the ribonucleotide C2'-position. (C) 2009 Elsevier Ltd. All rights reserved.
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