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• W3010348650 endingPage "3677" @default.
• W3010348650 startingPage "3657" @default.
• W3010348650 abstract "Abstract DNA replication is a central process in all living organisms. Polyomavirus DNA replication serves as a model system for eukaryotic DNA replication and has considerably contributed to our understanding of basic replication mechanisms. However, the details of the involved processes are still unclear, in particular regarding lagging strand synthesis. To delineate the complex mechanism of coordination of various cellular proteins binding simultaneously or consecutively to DNA to initiate replication, we investigated single-stranded DNA (ssDNA) interactions by the SV40 large T antigen (Tag). Using single molecule imaging by atomic force microscopy (AFM) combined with biochemical and spectroscopic analyses we reveal independent activity of monomeric and oligomeric Tag in high affinity binding to ssDNA. Depending on ssDNA length, we obtain dissociation constants for Tag-ssDNA interactions (KD values of 10–30 nM) that are in the same order of magnitude as ssDNA binding by human replication protein A (RPA). Furthermore, we observe the formation of RPA-Tag-ssDNA complexes containing hexameric as well as monomeric Tag forms. Importantly, our data clearly show stimulation of primase function in lagging strand Okazaki fragment synthesis by monomeric Tag whereas hexameric Tag inhibits the reaction, redefining DNA replication initiation on the lagging strand." @default.
• W3010348650 created "2020-03-13" @default.
• W3010348650 creator A5026180927 @default.
• W3010348650 creator A5039095695 @default.
• W3010348650 creator A5045600249 @default.
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• W3010348650 creator A5079270439 @default.
• W3010348650 creator A5081949907 @default.
• W3010348650 creator A5090427264 @default.
• W3010348650 date "2020-03-04" @default.
• W3010348650 modified "2023-10-03" @default.
• W3010348650 title "SV40 T antigen interactions with ssDNA and replication protein A: a regulatory role of T antigen monomers in lagging strand DNA replication" @default.
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