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Download Mario Is Missing Peach Untold Tale 2 0 2 >> Download.Absence of p300 enhances SV40 minichromosome maintenance replication in cell-free extracts.
To investigate the functions of p300 in replication of plasmids that replicate by a recombination-dependent mechanism, we have studied the replication of single-stranded minichromosomes containing the SV40 origin of replication in an in vitro transcription/translation system containing p300 and DNA-dependent protein kinase activity. Replication of such minichromosomes in this system is highly accurate and is blocked by the non-nucleoside inhibitor of DNA-dependent protein kinase, geldanamycin. In this report, we show that the presence of p300 negatively regulates the replication of such minichromosomes by complexing with Mr = 317,000 protein(s) and interacting with the C-terminal domain of DNA-dependent protein kinase, resulting in a decrease in the kinase activity of the enzyme. The presence of p300 also affected DNA synthesis by the large T antigen. Our results show that the presence of a limiting amount of p300 leads to a marked decrease in the amount of DNA synthesized. The effects of p300 appear to be directed specifically towards minichromosome replication. By substituting with p300 the small ATP-binding proteins found in HeLa cell extracts, we show that p300 is present in large molecular weight complexes, which interact with DNA polymerase delta. The recruitment of these large molecular weight complexes at the SV40 origin of replication is accompanied by an increase in their kin
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