PMID: 9544702Apr 17, 1998Paper

Roles of TraI protein with activities of cleaving and rejoining the single-stranded DNA in both initiation and termination of conjugal DNA transfer

Genes to Cells : Devoted to Molecular & Cellular Mechanisms
H Fukuda, E Ohtsubo

Abstract

The plasmid R100 encodes the TraI protein, which is required for conjugal DNA transfer. TraI has the activity of site- and strand-specific nicking of the supercoiled plasmid DNA. The molecular mechanism of this specific nicking, which is supposed to be the initiation reaction of DNA transfer, is not understood. We have demonstrated that TraI has the ability to cleave the single-stranded DNA at the same site as the nicking site (nic) in a region, which we here refer to as sbi. The product contained the TraI protein which was covalently linked to the newly generated 5' end of the nicking reaction. Both the cleaving and nicking reactions took place under almost the same conditions and required the presence of the sbi region. DNase I-footprinting analysis revealed that the TraI bound to the single-stranded DNA of the sbi region. TraI did not cleave the double-stranded DNA fragment, but it did cleave the double-stranded DNA with a single-stranded DNA portion in the sbi region. KMnO4 mapping analysis revealed that TraI can melt the sbi region in the supercoiled DNA to generate a single-stranded portion. We have also demonstrated that TraI was able to rejoin the cleaved products. The rejoining reaction required the 5' end of one cleav...Continue Reading

Citations

Jun 6, 2014·Journal of Molecular Modeling·Nicholas J ClarkJoseph E Curtis
Sep 25, 2003·Proceedings of the National Academy of Sciences of the United States of America·Matthew J Harley, Joel F Schildbach
Sep 12, 1998·Genes to Cells : Devoted to Molecular & Cellular Mechanisms·K TakiE Ohtsubo
Mar 31, 2018·Frontiers in Molecular Biosciences·Fabián Lorenzo-DíazManuel Espinosa
Mar 15, 2003·Biochimica Et Biophysica Acta·Lara M StreetJoel F Schildbach

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