In vitro Type II Restriction of Bacteriophage DNA With Modified Pyrimidines

Frontiers in Microbiology
Kiersten FlodmanShuang-Yong Xu

Abstract

To counteract host-encoded restriction systems, bacteriophages (phages) incorporate modified bases in their genomes. For example, phages carry in their genomes modified pyrimidines such as 5-hydroxymethyl-cytosine (5hmC) in T4gt deficient in α- and β-glycosyltransferases, glucosylated-5-hydroxymethylcytosine (5gmC) in T4, 5-methylcytosine (5mC) in Xp12, and 5-hydroxymethyldeoxyuridine (5hmdU) in SP8. In this work we sequenced phage Xp12 and SP8 genomes and examined Type II restriction of T4gt, T4, Xp12, and SP8 phage DNAs. T4gt, T4, and Xp12 genomes showed resistance to 81.9% (186 out of 227 enzymes tested), 94.3% (214 out of 227 enzymes tested), and 89.9% (196 out of 218 enzymes tested), respectively, commercially available Type II restriction endonucleases (REases). The SP8 genome, however, was resistant to only ∼8.3% of these enzymes (17 out of 204 enzymes tested). SP8 DNA could be further modified by adenine DNA methyltransferases (MTases) such as M.Dam and M.EcoGII as well as a number of cytosine DNA MTases, such as CpG methylase. The 5hmdU base in SP8 DNA was phosphorylated by treatment with a 5hmdU DNA kinase to achieve ∼20% phosphorylated 5hmdU, resulting resistance or partially resistant to more Type II restriction. Th...Continue Reading

References

Sep 1, 1976·Proceedings of the National Academy of Sciences of the United States of America·L SnyderE Kutter
Jan 1, 1992·Annual Review of Microbiology·P P HoetC G Cocito
Mar 1, 1969·Proceedings of the National Academy of Sciences of the United States of America·L M Gold, M Schweiger
Oct 1, 1968·Canadian Journal of Microbiology·T T KuoC P Chen
Mar 11, 1982·Nucleic Acids Research·L H HuangM Ehrlich
Sep 15, 2001·Nucleic Acids Research·A Pingoud, A Jeltsch
Mar 11, 2003·Microbiology and Molecular Biology Reviews : MMBR·Eric S MillerWolfgang Rüger
Mar 26, 2003·Nucleic Acids Research·Richard J RobertsShuang-yong Xu
Jun 26, 2003·Nucleic Acids Research·Tamas VinczeRichard J Roberts
Feb 12, 2008·BMC Genomics·Ramy K AzizOlga Zagnitko
Nov 22, 2011·Nucleic Acids Research·Marek DrozdzMonika Radlinska
Aug 31, 2013·Nucleic Acids Research·Wil A M Loenen, Elisabeth A Raleigh
May 23, 2014·Angewandte Chemie·Zuzana Vaníková, Michal Hocek
Nov 8, 2014·Nucleic Acids Research·Richard J RobertsDana Macelis
May 24, 2016·Journal of the American Chemical Society·Angad P MehtaPeter G Schultz
Jun 9, 2016·Nucleic Acids Research·Alfred PingoudWolfgang Wende
Jun 21, 2016·Chemical Reviews·Peter Weigele, Elisabeth A Raleigh
Mar 21, 2018·Proceedings of the National Academy of Sciences of the United States of America·Yan-Jiun LeePeter R Weigele
Apr 16, 2019·Frontiers in Microbiology·Kiersten FlodmanShuang-Yong Xu
Sep 11, 2019·Nucleic Acids Research·Thomas LutzShuang-Yong Xu

❮ Previous
Next ❯

Citations

Jun 23, 2021·Proceedings of the National Academy of Sciences of the United States of America·Evan J BurkeLana Saleh

❮ Previous
Next ❯

Datasets Mentioned

BETA
MT664984
MW001214

Methods Mentioned

BETA
Restriction Digestions
density gradient centrifugation
electrophoresis
PCR
restriction
in vitro transcription

Software Mentioned

Rapid Annotation of Subsystems Technology ( RAST )
NEBcutter

Related Concepts

Related Feeds

Bacteriophage: Phage Therapy

Phage therapy uses bacterial viruses (bacteriophages) to treat bacterial infections and is widely being recognized as an alternative to antibiotics. Here is the latest research.