Interaction between DNA polymerase λ and RPA during translesion synthesis

• Published in: Biochemistry (Moscow) 2008-09, Vol.73 (9), p.1042-1046
• Main Authors: Krasikova, Yu. S., Belousova, E. A., Lebedeva, N. A., Pestryakov, P. E., Lavrik, O. I.
• Format: Article
• Language: English
• Subjects: Animals; Biochemistry; Biomedical and Life Sciences; Biomedicine; Bioorganic Chemistry; DNA Damage; DNA Polymerase beta - metabolism; DNA Primers - genetics; DNA Primers - metabolism; DNA Replication; DNA, Bacterial - metabolism; Guanine - metabolism; Life Sciences; Microbiology; Replication Protein A - genetics; Replication Protein A - metabolism; Templates, Genetic
• ISSN: 0006-2979; 1608-3040
• DOI: 10.1134/S0006297908090125

Replication of damaged DNA (translesion synthesis, TLS) is realized by specialized DNA polymerases. Additional protein factors such as replication protein A (RPA) play important roles in this process. However, details of the interaction are unknown. Here we analyzed the influence of the hRPA and its mutant hABCD lacking domains responsible for protein-protein interactions on ability of DNA polymerase λ to catalyze TLS. The primer-template structures containing varying parts of extended strand (16 and 37 nt) were used as model systems imitating DNA intermediate of first stage of TLS. The 8-oxoguanine disposed in +1 position of the template strand in relation to 3′-end of primer was exploited as damage. It was shown that RPA stimulated TLS DNA synthesis catalyzed by DNA polymerase λ in its globular but not in extended conformation. Moreover, this effect is dependent on the presence of p70N and p32C domains in RPA molecule.