A double-strand break does not promote Neisseria gonorrhoeae pilin antigenic variation

Lauren L. Prister, Jing Xu, Hank Seifert*

*Corresponding author for this work

Research output: Contribution to journalArticle

Abstract

The major subunit of the type IV pilus (T4p) of Neisseria gonorrhoeae undergoes antigenic variation (AV) dependent on a guanine quadruplex (G4) DNA structure located upstream of the pilin gene. Since the presence of G4 DNA induces genome instability in both eukaryotic and prokaryotic chromosomes, we tested whether a double-strand break (DSB) at the site of the pilE G4 sequence could substitute for G4-directed pilin AV. The G4 motif was replaced by an I-SceI cut site, and the cut site was also introduced to locations near the origin of replication and the terminus. Expression of the I-SceI endonuclease from an irrelevant chromosomal site confirmed that the endonuclease functions to induce double-strand breaks at all three locations. No antigenic variants were detected when the G4 was replaced with the I-SceI cut site, but there was a growth defect from having a DSB in the chromosome, and suppressor mutations that were mainly deletions of the cut site and/or the entire pilE gene accumulated. Thus, the pilE G4 does not act to promote pilin AV by generating a DSB but requires either a different type of break, a nick, or more complex interactions with other factors to stimulate this programmed recombination system.

Original languageEnglish (US)
Article numbere00256-19
JournalJournal of bacteriology
Volume201
Issue number13
DOIs
StatePublished - Jan 1 2019

Fingerprint

Fimbriae Proteins
Antigenic Variation
Neisseria gonorrhoeae
G-Quadruplexes
Genetic Suppression
Chromosome Breakage
Replication Origin
Genomic Instability
Endonucleases
Deoxyribonuclease I
Genetic Recombination
Genes
Chromosomes
DNA
Growth

Keywords

  • Antigenic variation
  • Double-strand break
  • Guanine quadruplex
  • Homologous recombination
  • Pilus
  • RecA

ASJC Scopus subject areas

  • Microbiology
  • Molecular Biology

Cite this

@article{fe94cf0539884553b3f9d80deace3cd5,
title = "A double-strand break does not promote Neisseria gonorrhoeae pilin antigenic variation",
abstract = "The major subunit of the type IV pilus (T4p) of Neisseria gonorrhoeae undergoes antigenic variation (AV) dependent on a guanine quadruplex (G4) DNA structure located upstream of the pilin gene. Since the presence of G4 DNA induces genome instability in both eukaryotic and prokaryotic chromosomes, we tested whether a double-strand break (DSB) at the site of the pilE G4 sequence could substitute for G4-directed pilin AV. The G4 motif was replaced by an I-SceI cut site, and the cut site was also introduced to locations near the origin of replication and the terminus. Expression of the I-SceI endonuclease from an irrelevant chromosomal site confirmed that the endonuclease functions to induce double-strand breaks at all three locations. No antigenic variants were detected when the G4 was replaced with the I-SceI cut site, but there was a growth defect from having a DSB in the chromosome, and suppressor mutations that were mainly deletions of the cut site and/or the entire pilE gene accumulated. Thus, the pilE G4 does not act to promote pilin AV by generating a DSB but requires either a different type of break, a nick, or more complex interactions with other factors to stimulate this programmed recombination system.",
keywords = "Antigenic variation, Double-strand break, Guanine quadruplex, Homologous recombination, Pilus, RecA",
author = "Prister, {Lauren L.} and Jing Xu and Hank Seifert",
year = "2019",
month = "1",
day = "1",
doi = "10.1128/JB.00256-19",
language = "English (US)",
volume = "201",
journal = "Journal of Bacteriology",
issn = "0021-9193",
publisher = "American Society for Microbiology",
number = "13",

}

A double-strand break does not promote Neisseria gonorrhoeae pilin antigenic variation. / Prister, Lauren L.; Xu, Jing; Seifert, Hank.

In: Journal of bacteriology, Vol. 201, No. 13, e00256-19, 01.01.2019.

Research output: Contribution to journalArticle

TY - JOUR

T1 - A double-strand break does not promote Neisseria gonorrhoeae pilin antigenic variation

AU - Prister, Lauren L.

AU - Xu, Jing

AU - Seifert, Hank

PY - 2019/1/1

Y1 - 2019/1/1

N2 - The major subunit of the type IV pilus (T4p) of Neisseria gonorrhoeae undergoes antigenic variation (AV) dependent on a guanine quadruplex (G4) DNA structure located upstream of the pilin gene. Since the presence of G4 DNA induces genome instability in both eukaryotic and prokaryotic chromosomes, we tested whether a double-strand break (DSB) at the site of the pilE G4 sequence could substitute for G4-directed pilin AV. The G4 motif was replaced by an I-SceI cut site, and the cut site was also introduced to locations near the origin of replication and the terminus. Expression of the I-SceI endonuclease from an irrelevant chromosomal site confirmed that the endonuclease functions to induce double-strand breaks at all three locations. No antigenic variants were detected when the G4 was replaced with the I-SceI cut site, but there was a growth defect from having a DSB in the chromosome, and suppressor mutations that were mainly deletions of the cut site and/or the entire pilE gene accumulated. Thus, the pilE G4 does not act to promote pilin AV by generating a DSB but requires either a different type of break, a nick, or more complex interactions with other factors to stimulate this programmed recombination system.

AB - The major subunit of the type IV pilus (T4p) of Neisseria gonorrhoeae undergoes antigenic variation (AV) dependent on a guanine quadruplex (G4) DNA structure located upstream of the pilin gene. Since the presence of G4 DNA induces genome instability in both eukaryotic and prokaryotic chromosomes, we tested whether a double-strand break (DSB) at the site of the pilE G4 sequence could substitute for G4-directed pilin AV. The G4 motif was replaced by an I-SceI cut site, and the cut site was also introduced to locations near the origin of replication and the terminus. Expression of the I-SceI endonuclease from an irrelevant chromosomal site confirmed that the endonuclease functions to induce double-strand breaks at all three locations. No antigenic variants were detected when the G4 was replaced with the I-SceI cut site, but there was a growth defect from having a DSB in the chromosome, and suppressor mutations that were mainly deletions of the cut site and/or the entire pilE gene accumulated. Thus, the pilE G4 does not act to promote pilin AV by generating a DSB but requires either a different type of break, a nick, or more complex interactions with other factors to stimulate this programmed recombination system.

KW - Antigenic variation

KW - Double-strand break

KW - Guanine quadruplex

KW - Homologous recombination

KW - Pilus

KW - RecA

UR - http://www.scopus.com/inward/record.url?scp=85067909470&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=85067909470&partnerID=8YFLogxK

U2 - 10.1128/JB.00256-19

DO - 10.1128/JB.00256-19

M3 - Article

C2 - 30988037

AN - SCOPUS:85067909470

VL - 201

JO - Journal of Bacteriology

JF - Journal of Bacteriology

SN - 0021-9193

IS - 13

M1 - e00256-19

ER -