Pol31 and Pol32 subunits of yeast DNA polymerase δ are also essential subunits of DNA polymerase ζ

Abstract

Replication through a diverse array of DNA lesions occurs by the sequential action of two translesion synthesis (TLS) DNA polymerases (Pols), in which one inserts the nucleotide opposite the lesion and the other carries out the subsequent extension. By extending from the nucleotide inserted by another Pol, Polζ plays an indispensable role in mediating lesion bypass. Polζ comprises the Rev3 catalytic and Rev7 accessory subunits. Pol32, a subunit of the replicative polymerase Polδ, is also required for Polζ-dependent TLS, but how this Polδ subunit contributes to Polζ function in TLS has remained unknown. Here we show that yeast Polζ is a four-subunit enzyme containing Rev3, Rev7, Pol31, and Pol32; in this complex, association with Pol31/Pol32 is mediated via binding of the Rev3 C terminus to Pol31. The functional requirement of this complex is supported by evidence that mutational inactivation of Rev3's ability to bind Pol31 abrogates Polζ's role in TLS in yeast cells. These findings identify an unexpected role of Pol31 and Pol32 as two essential subunits of Polζ, and clarify why these proteins are required for Polζ-dependent TLS, but not for TLS mediated by Polη in yeast cells. To distinguish the four-subunit complex from the two-subunit Polζ, we designate the four-subunit enzyme "Polζ-d," where "-d" denotes the Pol31/Pol32 subunits of Polδ.