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 language | English (US) |
|---|---|
| Article number | e00256-19 |
| Journal | Journal of bacteriology |
| Volume | 201 |
| Issue number | 13 |
| DOIs | |
| State | Published - Jan 1 2019 |
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Keywords
- Antigenic variation
- Double-strand break
- Guanine quadruplex
- Homologous recombination
- Pilus
- RecA
ASJC Scopus subject areas
- Microbiology
- Molecular Biology
Cite this
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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 journal › Article
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 -