Methylomic and phenotypic analysis of the ModH5 phasevarion of Helicobacter pylori

Yogitha N. Srikhanta, Rebecca J. Gorrell, Peter Power, Kirill Tsyganov, Matthew Boitano, Tyson A Clark, Jonas Korlach, Elizabeth L. Hartland, Michael P Jennings, Terry Kwok

Research output: Contribution to journalArticleResearchpeer-review

Abstract

The Helicobacter pylori phase variable gene modH, typified by gene HP1522 in strain 26695, encodes a N6-adenosine type III DNA methyltransferase. Our previous studies identified multiple strain-specific modH variants (modH1 - modH19) and showed that phase variation of modH5 in H. pylori P12 influenced expression of motility-associated genes and outer membrane protein gene hopG. However, the ModH5 DNA recognition motif and the mechanism by which ModH5 controls gene expression were unknown. Here, using comparative single molecule real-time sequencing, we identify the DNA site methylated by ModH5 as 5′-Gm6ACC-3′. This motif is vastly underrepresented in H. pylori genomes, but overrepresented in a number of virulence genes, including motility-associated genes, and outer membrane protein genes. Motility and the number of flagella of H. pylori P12 wild-type were significantly higher than that of isogenic modH5 OFF or ΔmodH5 mutants, indicating that phase variable switching of modH5 expression plays a role in regulating H. pylori motility phenotypes. Using the flagellin A (flaA) gene as a model, we show that ModH5 modulates flaA promoter activity in a GACC methylation-dependent manner. These findings provide novel insights into the role of ModH5 in gene regulation and how it mediates epigenetic regulation of H. pylori motility.

Original languageEnglish
Article number16140
Number of pages14
JournalScientific Reports
Volume7
Issue number1
DOIs
Publication statusPublished - 1 Dec 2017

Keywords

  • bacterial genetics
  • DNA methylation
  • infection

Cite this

Srikhanta, Yogitha N. ; Gorrell, Rebecca J. ; Power, Peter ; Tsyganov, Kirill ; Boitano, Matthew ; Clark, Tyson A ; Korlach, Jonas ; Hartland, Elizabeth L. ; Jennings, Michael P ; Kwok, Terry. / Methylomic and phenotypic analysis of the ModH5 phasevarion of Helicobacter pylori. In: Scientific Reports. 2017 ; Vol. 7, No. 1.
@article{17dcd185305c4cdb8039948fbc31379c,
title = "Methylomic and phenotypic analysis of the ModH5 phasevarion of Helicobacter pylori",
abstract = "The Helicobacter pylori phase variable gene modH, typified by gene HP1522 in strain 26695, encodes a N6-adenosine type III DNA methyltransferase. Our previous studies identified multiple strain-specific modH variants (modH1 - modH19) and showed that phase variation of modH5 in H. pylori P12 influenced expression of motility-associated genes and outer membrane protein gene hopG. However, the ModH5 DNA recognition motif and the mechanism by which ModH5 controls gene expression were unknown. Here, using comparative single molecule real-time sequencing, we identify the DNA site methylated by ModH5 as 5′-Gm6ACC-3′. This motif is vastly underrepresented in H. pylori genomes, but overrepresented in a number of virulence genes, including motility-associated genes, and outer membrane protein genes. Motility and the number of flagella of H. pylori P12 wild-type were significantly higher than that of isogenic modH5 OFF or ΔmodH5 mutants, indicating that phase variable switching of modH5 expression plays a role in regulating H. pylori motility phenotypes. Using the flagellin A (flaA) gene as a model, we show that ModH5 modulates flaA promoter activity in a GACC methylation-dependent manner. These findings provide novel insights into the role of ModH5 in gene regulation and how it mediates epigenetic regulation of H. pylori motility.",
keywords = "bacterial genetics, DNA methylation, infection",
author = "Srikhanta, {Yogitha N.} and Gorrell, {Rebecca J.} and Peter Power and Kirill Tsyganov and Matthew Boitano and Clark, {Tyson A} and Jonas Korlach and Hartland, {Elizabeth L.} and Jennings, {Michael P} and Terry Kwok",
year = "2017",
month = "12",
day = "1",
doi = "10.1038/s41598-017-15721-x",
language = "English",
volume = "7",
journal = "Scientific Reports",
issn = "2045-2322",
publisher = "Nature Publishing Group",
number = "1",

}

Methylomic and phenotypic analysis of the ModH5 phasevarion of Helicobacter pylori. / Srikhanta, Yogitha N.; Gorrell, Rebecca J.; Power, Peter; Tsyganov, Kirill; Boitano, Matthew; Clark, Tyson A; Korlach, Jonas; Hartland, Elizabeth L.; Jennings, Michael P; Kwok, Terry.

In: Scientific Reports, Vol. 7, No. 1, 16140, 01.12.2017.

Research output: Contribution to journalArticleResearchpeer-review

TY - JOUR

T1 - Methylomic and phenotypic analysis of the ModH5 phasevarion of Helicobacter pylori

AU - Srikhanta, Yogitha N.

AU - Gorrell, Rebecca J.

AU - Power, Peter

AU - Tsyganov, Kirill

AU - Boitano, Matthew

AU - Clark, Tyson A

AU - Korlach, Jonas

AU - Hartland, Elizabeth L.

AU - Jennings, Michael P

AU - Kwok, Terry

PY - 2017/12/1

Y1 - 2017/12/1

N2 - The Helicobacter pylori phase variable gene modH, typified by gene HP1522 in strain 26695, encodes a N6-adenosine type III DNA methyltransferase. Our previous studies identified multiple strain-specific modH variants (modH1 - modH19) and showed that phase variation of modH5 in H. pylori P12 influenced expression of motility-associated genes and outer membrane protein gene hopG. However, the ModH5 DNA recognition motif and the mechanism by which ModH5 controls gene expression were unknown. Here, using comparative single molecule real-time sequencing, we identify the DNA site methylated by ModH5 as 5′-Gm6ACC-3′. This motif is vastly underrepresented in H. pylori genomes, but overrepresented in a number of virulence genes, including motility-associated genes, and outer membrane protein genes. Motility and the number of flagella of H. pylori P12 wild-type were significantly higher than that of isogenic modH5 OFF or ΔmodH5 mutants, indicating that phase variable switching of modH5 expression plays a role in regulating H. pylori motility phenotypes. Using the flagellin A (flaA) gene as a model, we show that ModH5 modulates flaA promoter activity in a GACC methylation-dependent manner. These findings provide novel insights into the role of ModH5 in gene regulation and how it mediates epigenetic regulation of H. pylori motility.

AB - The Helicobacter pylori phase variable gene modH, typified by gene HP1522 in strain 26695, encodes a N6-adenosine type III DNA methyltransferase. Our previous studies identified multiple strain-specific modH variants (modH1 - modH19) and showed that phase variation of modH5 in H. pylori P12 influenced expression of motility-associated genes and outer membrane protein gene hopG. However, the ModH5 DNA recognition motif and the mechanism by which ModH5 controls gene expression were unknown. Here, using comparative single molecule real-time sequencing, we identify the DNA site methylated by ModH5 as 5′-Gm6ACC-3′. This motif is vastly underrepresented in H. pylori genomes, but overrepresented in a number of virulence genes, including motility-associated genes, and outer membrane protein genes. Motility and the number of flagella of H. pylori P12 wild-type were significantly higher than that of isogenic modH5 OFF or ΔmodH5 mutants, indicating that phase variable switching of modH5 expression plays a role in regulating H. pylori motility phenotypes. Using the flagellin A (flaA) gene as a model, we show that ModH5 modulates flaA promoter activity in a GACC methylation-dependent manner. These findings provide novel insights into the role of ModH5 in gene regulation and how it mediates epigenetic regulation of H. pylori motility.

KW - bacterial genetics

KW - DNA methylation

KW - infection

UR - http://www.scopus.com/inward/record.url?scp=85034850441&partnerID=8YFLogxK

U2 - 10.1038/s41598-017-15721-x

DO - 10.1038/s41598-017-15721-x

M3 - Article

VL - 7

JO - Scientific Reports

JF - Scientific Reports

SN - 2045-2322

IS - 1

M1 - 16140

ER -