Nucleic Acids Research Advance Access originally published online on August 26, 2006
Nucleic Acids Research 2006 34(16):4335-4341; doi:10.1093/nar/gkl403
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Nucleic Acids Research, 2006, Vol. 34, No. 16 4335-4341
© 2006 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 |
RPA and PCNA suppress formation of large deletion errors by yeast DNA polymerase 
1 Laboratory of Molecular Genetics, National Institute of Environmental Health Sciences, NIH, DHHS Research Triangle Park, NC 27709, USA 2 Laboratory of Structural Biology, National Institute of Environmental Health Sciences, NIH, DHHS Research Triangle Park, NC 27709, USA 3 Biostatistics Branch, National Institute of Environmental Health Sciences, NIH, DHHS Research Triangle Park, NC 27709, USA 4 Department of Biochemistry and Molecular Biophysics, Washington University St Louis, MO 63110, USA
*To whom correspondence should be addressed. Tel: +1 919 541 2644; Fax: +1 919 541 7613; Email: kunkel{at}niehs.nih.gov
Received April 10, 2006. Revised May 11, 2006. Accepted May 14, 2006.
In fulfilling its biosynthetic roles in nuclear replication and in several types of repair, DNA polymerase 
(pol ) is assisted by replication protein A (RPA), the single-stranded DNA-binding protein complex, and by the processivity clamp proliferating cell nuclear antigen (PCNA). Here we report the effects of these accessory proteins on the fidelity of DNA synthesis in vitro by yeast pol . We show that when RPA and PCNA are included in reactions containing pol , rates for single base errors are similar to those generated by pol alone, indicating that pol itself is by far the prime determinant of fidelity for single base errors. However, the rate of deleting multiple nucleotides between directly repeated sequences is reduced by 
10-fold in the presence of either RPA or PCNA, and by 
90-fold when both proteins are present. We suggest that PCNA and RPA suppress large deletion errors by preventing the primer terminus at a repeat from fraying and/or from relocating and annealing to a downstream repeat. Strong suppression of deletions by PCNA and RPA suggests that they may contribute to the high replication fidelity needed to stably maintain eukaryotic genomes that contain abundant repetitive sequences.
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