An oligonucleotide inhibits oligomerization of a rolling circle initiator protein at the pT181 origin of replication.

The Journal of Biological Chemistry
A C ZhaoS A Khan

Abstract

A large number of plasmids have been shown to replicate by a rolling circle (RC) mechanism. The initiators encoded by these plasmids have origin-specific, nicking-closing activity that is required for the initiation and termination of RC replication. Since the initiators of many RC plasmids are rate-limiting for replication, these proteins are usually inactivated after supporting one round of replication. In the case of the pT181 plasmid, inactivation of the initiator RepC protein occurs by the attachment of an oligonucleotide to its active tyrosine residue. We have generated the inactivated form of RepC, termed RepC*, in vitro and investigated the effects of attachment of the oligonucleotide on its various biochemical activities. Our results demonstrate that while RepC* is inactive in nicking-closing and replication activities due to the blockage of its active tyrosine residue, it is competent in origin DNA binding and DNA religation activities. We have investigated the oligomeric state of RepC and RepC* and found that RepC exists as a dimer in solution and can oligomerize on the DNA. We have generated heterodimers in vitro between the wild-type and epitope-tagged RepC proteins. In electrophoretic mobility shift experiments, t...Continue Reading

References

Apr 1, 1992·Proceedings of the National Academy of Sciences of the United States of America·L A DempseyS A Khan
Nov 1, 1990·Proceedings of the National Academy of Sciences of the United States of America·P NoirotR P Novick
Jun 1, 1989·Microbiological Reviews·A Gruss, S D Ehrlich
Jun 1, 1985·Microbiological Reviews·N D Zinder, K Horiuchi
Oct 1, 1985·Proceedings of the National Academy of Sciences of the United States of America·R R KoepselS A Khan
Aug 1, 1986·Proceedings of the National Academy of Sciences of the United States of America·R R KoepselS A Khan

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Citations

Jan 14, 2020·Nucleic Acids Research·Carolina CarrascoFernando Moreno-Herrero
Aug 10, 2000·Molecular Microbiology·S A Khan
Jun 25, 2015·Microbiology Spectrum·J A Ruiz-MasóG Del Solar

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