Journal
MOLECULAR AND CELLULAR BIOLOGY
Volume 20, Issue 19, Pages 7099-7108Publisher
AMER SOC MICROBIOLOGY
DOI: 10.1128/MCB.20.19.7099-7108.2000
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DNA polymerase activity is essential for replication, recombination, repair, and mutagenesis. All DNA polymerases studied so far from any biological source synthesize DNA by the Watson-Crick base-pairing rule, incorporating A, G, C, and T opposite the templates T, C, G, and A, respectively. Non-Watson-Crick base pairs would lead to mutations. In this report, we describe the ninth human DNA polymerase, Pol iota, encoded by the RAD30B gene. We show that human Pol iota violates the Watson-Crick base-pairing rule opposite template T. During base selection, human Pol iota preferred T-G base pairing, leading to G incorporation opposite template T. The resulting T-G base pair was less efficiently extended by human Pol iota compared to the Watson-Crick base pairs. Consequently, DNA synthesis frequently aborted opposite template T, a property we designated the T stop. This T stop restricted human Pol iota to a very short stretch of DNA synthesis. Furthermore, kinetic analyses show that human Pul iota copies template C with extraordinarily low fidelity, misincorporating T, A, and C with unprecedented frequencies of 1/9, 1/10, and 1/11, respectively. Human Pol iota incorporated one nucleotide opposite a template abasic site more efficiently than opposite a template T, suggesting a role for human Pol iota in DNA lesion bypass. The unique features of preferential G incorporation opposite template T and T stop suggest that DNA Pull may additionally play a specialized function in human biology.
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