DpnA, a methylase for single-strand DNA in the Dpn II restriction system, and its biological function
Abstract
The two DNA-adenine methylases encoded by the Dpn II restriction gene cassette were purified, and their activities were compared on various DNA substrates. DpnA was able to methylate single-strand DNA and double-strand DNA, whereas DpnM methylated only double-strand DNA. Although both enzymes act at 5{prime}-GATC-3{prime} in DNA, DpnA can also methylate sequences altered in the guanine position, but at a lower rate. A deletion mutation in the dpnA gene was constructed and transferred to the chromosome. Transmission by way of the transformation pathway of methylated and unmethylated plasmids to dpnA mutant and wild-type recipients was examined. The mutant cells restricted unmethylated donor plasmic establishment much more strongly than did wild-type cells. In the wild type, the single strands of donor plasmid DNA that enter by the transformation pathway are apparently methylated by DpnA prior to conversion of the plasmid to a double-strand form, in which the plasmid would be susceptible to the Dpn II endonuclease. The biological function of DpnA may, therefore, be the enhancement of plasmid transfer to Dpn II-containing strains of Streptococcus pneumoniae.
- Authors:
-
- Brookhaven National Lab., Upton, NY (USA)
- Publication Date:
- OSTI Identifier:
- 7138316
- Resource Type:
- Journal Article
- Journal Name:
- Proceedings of the National Academy of Sciences of the United States of America; (USA)
- Additional Journal Information:
- Journal Volume: 86:23; Journal Issue: 23; Journal ID: ISSN 0027-8424
- Country of Publication:
- United States
- Language:
- English
- Subject:
- 59 BASIC BIOLOGICAL SCIENCES; ESCHERICHIA COLI; CELL TRANSFORMATIONS; HYDROLASES; GENE MUTATIONS; PLASMIDS; GENE RECOMBINATION; STREPTOCOCCUS; ADENINES; DNA SEQUENCING; GENES; OLIGONUCLEOTIDES; AMINES; ANTIMETABOLITES; AROMATICS; AZAARENES; BACTERIA; CELL CONSTITUENTS; DRUGS; ENZYMES; HETEROCYCLIC COMPOUNDS; MICROORGANISMS; MUTATIONS; NUCLEIC ACIDS; ORGANIC COMPOUNDS; ORGANIC NITROGEN COMPOUNDS; PURINES; STRUCTURAL CHEMICAL ANALYSIS; 550200* - Biochemistry
Citation Formats
Cerritelli, S, Springhorn, S S, and Lacks, S A. DpnA, a methylase for single-strand DNA in the Dpn II restriction system, and its biological function. United States: N. p., 1989.
Web. doi:10.1073/pnas.86.23.9223.
Cerritelli, S, Springhorn, S S, & Lacks, S A. DpnA, a methylase for single-strand DNA in the Dpn II restriction system, and its biological function. United States. https://doi.org/10.1073/pnas.86.23.9223
Cerritelli, S, Springhorn, S S, and Lacks, S A. 1989.
"DpnA, a methylase for single-strand DNA in the Dpn II restriction system, and its biological function". United States. https://doi.org/10.1073/pnas.86.23.9223.
@article{osti_7138316,
title = {DpnA, a methylase for single-strand DNA in the Dpn II restriction system, and its biological function},
author = {Cerritelli, S and Springhorn, S S and Lacks, S A},
abstractNote = {The two DNA-adenine methylases encoded by the Dpn II restriction gene cassette were purified, and their activities were compared on various DNA substrates. DpnA was able to methylate single-strand DNA and double-strand DNA, whereas DpnM methylated only double-strand DNA. Although both enzymes act at 5{prime}-GATC-3{prime} in DNA, DpnA can also methylate sequences altered in the guanine position, but at a lower rate. A deletion mutation in the dpnA gene was constructed and transferred to the chromosome. Transmission by way of the transformation pathway of methylated and unmethylated plasmids to dpnA mutant and wild-type recipients was examined. The mutant cells restricted unmethylated donor plasmic establishment much more strongly than did wild-type cells. In the wild type, the single strands of donor plasmid DNA that enter by the transformation pathway are apparently methylated by DpnA prior to conversion of the plasmid to a double-strand form, in which the plasmid would be susceptible to the Dpn II endonuclease. The biological function of DpnA may, therefore, be the enhancement of plasmid transfer to Dpn II-containing strains of Streptococcus pneumoniae.},
doi = {10.1073/pnas.86.23.9223},
url = {https://www.osti.gov/biblio/7138316},
journal = {Proceedings of the National Academy of Sciences of the United States of America; (USA)},
issn = {0027-8424},
number = 23,
volume = 86:23,
place = {United States},
year = {Fri Dec 01 00:00:00 EST 1989},
month = {Fri Dec 01 00:00:00 EST 1989}
}