RPA and PCNA suppress formation of large deletion errors by yeast DNA polymerase delta.


In fulfilling its biosynthetic roles in nuclear replication and in ...
In fulfilling its biosynthetic roles in nuclear replication and in several types of repair, DNA polymerase delta (pol delta) 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 delta. We show that when RPA and PCNA are included in reactions containing pol delta, rates for single base errors are similar to those generated by pol delta alone, indicating that pol delta 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 approximately 10-fold in the presence of either RPA or PCNA, and by > or =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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