Interaction of hREV1 with three human Y‐family DNA polymerases

Abstract

Polκ κ κ κ is one of many DNA polymerases involved in translesion DNA synthesis (TLS). It belongs to the Y-family of polymerases along with Polη η η η, Polι ι ι ι and hREV1. Unlike Polη η η η encoded by the xeroderma pigmentosum variant (XPV ) gene, Polκ κ κ κ is unable to bypass UV-induced DNA damage in vitro, but it is able to bypass benzo[a]pyrene (B[a]P)-adducted guanines accurately and efficiently. In an attempt to identify factor(s) targeting Polκ κ κ κ to its cognate DNA lesion(s), we searched for Polκ κ κ κ-interacting proteins by using the yeast two-hybrid assay. We found that Polκ κ κ κ interacts with a C-terminal region of hREV1. Polη η η η and Polι ι ι ι were also found to interact with the same region of hREV1. The interaction between Polκ κ κ κ and hREV1 was confirmed by pull-down and coimmunoprecipitation assays. The C-terminal region of hREV1 is known to interact with hREV7, a non-catalytic subunit of Polζ ζ ζ ζ that is another structurally unrelated TLS enzyme, and we show that Polκ κ κ κ and hREV7 bind to the same C-terminal region of hREV1. Thus, our results suggest that hREV1 plays a pivotal role in the multi-enzyme, multi-step process of translesion DNA synthesis.

The Vital Role of Polymerase ζ and REV1 in Mutagenic, but Not Correct, DNA Synthesis across Benzo[a]pyrene-dG and Recruitment of Polymerase ζ by REV1 to Replication-stalled Site

“…We show that the REV1 C terminus is important to the pol -catalyzed error-prone TLS ( Table 1). The same region of REV1 interacts with the other Y family polymerases (13,14), but its biological significance is unclear at present. The importance of the C terminus of REV1 protein to DNA damage tolerance has been reported in yeast (42) and chicken DT40 cells (26).…”

“…This enzyme activity plays a role in TLS across certain classes of lesions (10,11). REV1 also plays a noncatalytic role in TLS (12) and physically interacts with the other Y family polymerases (13,14) and the REV7 subunit of pol (15,16). The biological significance of these interactions remains to be determined.…”

Structural Basis of Recruitment of DNA Polymerase ζ by Interaction between REV1 and REV7 Proteins

“…Interestingly, REV7, a non-catalytic subunit of Pol, interacts with the C-terminal domain of REV1 ( Fig. 1) (7,9). This implies that REV1-REV7 interaction should exclude inserter polymerases from REV1 for polymerase switching to the extender polymerase.…”

Mouse models of DNA polymerases

“…The C-terminus of REV1 interacts with all the other Y family polymerases as well as the REV7 subunit of Pol f and PCNA [D'Souza et al, 2006]. Interestingly, these interactions do not affect the DNA polymerase activity or translesion synthesis ability of Rev1 [Guo et al, 2003;Ohashi et al, 2004]. Rev1 localizes to the nucleus.…”