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Report No.

${it Arabidopsis thaliana}$ Y-family DNA polymerase $$eta$$ catalyses translesion synthesis and interacts functionally with PCNA2

Anderson, H.*; Vonarx, E.*; Pastushok, L.*; Nakagawa, Mayu; Katafuchi, Atsushi*; Gruz, P.*; Rubbo, A.*; Grice, D.*; Osmond, M.*; Sakamoto, Ayako; Nomi, Takehiko*; Xiao, W.*; Kunz, B.*

We assessed the roles of ${it Arabidopsis thaliana}$ ${it POLH, PCNA1}$ and ${it PCNA2}$ in TLS mediated UV resistance. ${it Arabidopsis}$ ${it POLH}$ defective mutants sensitized growth of roots and whole plants to UV radiation indicating AtPol$$eta$$ contributes to UV resistance. ${it POLH}$ alone did not complement the UV sensitivity conferred by deletion of yeast ${it RAD30}$, although AtPol$$eta$$ exhibited cyclobutane dimer bypass activity and interacted with yeast PCNA in vitro. Co-expression of ${it POLH}$ and ${it PCNA2}$, but not ${it PCNA1}$, restored normal UV resistance and mutation kinetics in the ${it rad30}$ mutant. A single residue difference at site 201, which lies adjacent to the lysine ubiquitylated in PCNA, appeared responsible for the inability of PCNA1 to function with AtPol$$eta$$ in UV treated yeast. PCNA interacting protein boxes and an ubiquitin-binding motif in AtPol$$eta$$ were found to be required for restoration of UV resistance in the ${it rad30}$ mutant by ${it POLH}$ and ${it PCNA2}$. These observations indicate AtPol$$eta$$ can catalyse TLS past UV induced DNA damage, and link the biological activity of AtPol$$eta$$ in UV irradiated cells to PCNA2 and PCNA-and ubiquitin-binding motifs in AtPol$$eta$$.



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Category:Plant Sciences



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