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Human mitochondrial DNA contains more UV-induced lesions than the nuclear DNA due to lack of mechanism to remove bulky photoproducts. Human DNA polymerase gamma (Pol γ) is the sole DNA replicase in mitochondria, which contains a polymerase ( pol ) and an exonuclease ( exo ) active site. Previous studies showed that Pol γ only displays UV lesion bypassing when its exonuclease activity is obliterated. To investigate the reaction environment on Pol γ translesion activity, we tested Pol γ DNA activity in the presence of different metal ions. While Pol γ is unable to replicate through UV lesions on DNA templates in the presence of Mg 2+ , it exhibits robust translesion DNA synthesis (TLS) on cyclobutane pyrimidine dimer (CPD)-containing template when Mg 2+  was mixed with or completely replaced by Mn 2+ . Under these conditions, the efficiency of Pol γ′s TLS opposite CPD is near to that on a non-damaged template and is 800-fold higher than that of exonuclease-deficient Pol γ. Interestingly, Pol γ exhibits higher exonuclease activity in the presence of Mn 2+  than with Mg 2+ , suggesting Mn 2+ -stimulated Pol γ TLS is not via suppressing its exonuclease activity. We suggest that Mn 2+  ion expands Pol γ′s  pol  active site relative to Mg 2+  so that a UV lesion can be accommodated and blocks the communication between  pol  and  exo  active sites to execute translesion DNA synthesis.

Human Mitochondrial DNA Polymerase Metal Dependent UV Lesion Bypassing Ability