Sequence requirements for the termination of rolling-circle replication of plasmid pT181

Molecular Microbiology
A C Zhao, S A Khan

Abstract

Most small multicopy plasmids of Gram-positive bacteria and many in Gram-negative bacteria replicate by a rolling-circle (RC) mechanism. The replication initiator proteins encoded by the RC plasmids and single-stranded bacteriophages of Escherichia coli have origin-specific nicking-closing activities that are required for the initiation and termination of RC replication. We have investigated the sequence requirements for termination of RC replication of plasmid pT181. The initiator nick site is located in the loop of a hairpin region (IRII) within the pT181 origin of replication. By mutational analysis, we have found that several nucleotides within the stem of IRII which are critical for the initiation activity are dispensable for termination of replication. We also demonstrate that nucleotides in the right arm of IRII, but not the left arm, are absolutely required for termination of RC replication. We have also identified specific nucleotides in IRII that are critical for its termination activity. The sequence of the right arm of the hairpin must be located downstream of the initiator nick site for termination, suggesting that termination requires a specific orientation of the initiator protein at the origin.

Citations

Dec 19, 2017·Frontiers in Microbiology·Paweł WawrzyniakDariusz Bartosik
Jul 29, 2008·Homeopathy : the Journal of the Faculty of Homeopathy·V Bharatan
Feb 12, 2009·Nucleic Acids Research·Siu Kai Leung, Joseph T Y Wong
Aug 23, 2006·Molecular Systems Biology·Anthony C Forster, George M Church
Jun 10, 1998·Microbiology and Molecular Biology Reviews : MMBR·G del SolarR Díaz-Orejas
Dec 31, 1997·Microbiology and Molecular Biology Reviews : MMBR·S A Khan

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