Skip Navigation


Nucleic Acids Research Advance Access originally published online on March 12, 2009
Nucleic Acids Research 2009 37(9):2854-2866; doi:10.1093/nar/gkp155
This Article
Right arrow Full Text Freely available
Right arrow Print PDF (16025K) Freely available
Right arrow Screen PDF (785K) Freely available
Right arrow Supplementary Data
Right arrowOA All Versions of this Article:
37/9/2854    most recent
gkp155v1
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Services
Right arrow Email this article to a friend
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Add to My Personal Archive
Right arrow Download to citation manager
Right arrow Search for citing articles in:
ISI Web of Science (1)
Right arrow Commercial Re-use Guidelines
for Open Access NAR Content
Google Scholar
Right arrow Articles by Fazlieva, R.
Right arrow Articles by Matsumoto, Y.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Fazlieva, R.
Right arrow Articles by Matsumoto, Y.
Social Bookmarking
Add to CiteULike   Add to Connotea   Add to Del.icio.us  
What's this?

Nucleic Acids Research, 2009, Vol. 37, No. 9 2854-2866
© 2009 The Author(s)
This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/2.0/uk/) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.

Nucleic Acid Enzymes

Proofreading exonuclease activity of human DNA polymerase {delta} and its effects on lesion-bypass DNA synthesis

Ruzaliya Fazlieva1, Cynthia S. Spittle2, Darlene Morrissey1, Harutoshi Hayashi1, Hong Yan3 and Yoshihiro Matsumoto1,*

1Medical Science Division, 2Cancer Biomarker and Genotyping Facility and 3Institute for Cancer Research, Fox Chase Cancer Center, Philadelphia, PA 19111, USA

*To whom correspondence should be addressed. Tel: +1 215 728 5272; Fax: +1 215 728 4333; Email: yoshihiro.matsumoto{at}fccc.edu

Received February 19, 2009. Revised February 19, 2009. Accepted February 24, 2009.

Replicative DNA polymerases possess 3' -> 5' exonuclease activity to reduce misincorporation of incorrect nucleotides by proofreading during replication. To examine if this proofreading activity modulates DNA synthesis of damaged templates, we constructed a series of recombinant human DNA polymerase {delta} (Pol {delta}) in which one or two of the three conserved Asp residues in the exonuclease domain are mutated, and compared their properties with that of the wild-type enzyme. While all the mutant enzymes lost more than 95% exonuclease activity and severely decreased the proofreading activity than the wild-type, the bypass efficiency of damaged templates was varied: two mutant enzymes, D515V and D402A/D515A, gave higher bypass efficiencies on templates containing an abasic site, but another mutant, D316N/D515A, showed a lower bypass efficiency than the wild-type. All the enzymes including the wild-type inserted an adenine opposite the abasic site, whereas these enzymes inserted cytosine and adenine opposite an 8-oxoguanine with a ratio of 6:4. These results indicate that the exonuclease activity of human Pol {delta} modulates its intrinsic bypass efficiency on the damaged template, but does not affect the choice of nucleotide to be inserted.

Present Addresses: Cynthia S. Spittle, Wyeth, Collegeville, PA, USA.

Darlene Morrissey, St. Luke's Hospital, Bethlehem, PA, USA.

Harutoshi Hayashi, Haru Pets Clinic, Kasugai, Aichi, Japan.


Add to CiteULike CiteULike   Add to Connotea Connotea   Add to Del.icio.us Del.icio.us    What's this?




Disclaimer: Please note that abstracts for content published before 1996 were created through digital scanning and may therefore not exactly replicate the text of the original print issues. All efforts have been made to ensure accuracy, but the Publisher will not be held responsible for any remaining inaccuracies. If you require any further clarification, please contact our Customer Services Department.