The Neisseria gonorrhoeae photolyase orthologue phrB is required for proper DNA supercoiling but does not function in photo-reactivation.
Neisseria gonorrhoeae (Gc) is an obligate human pathogen and the causative agent of the sexually transmitted infection, gonorrhoea. Despite the fact that the gonococcus is not normally exposed to UV irradiation or visible light, the bacterium expresses a phrB orthologue, which in other organisms encodes a DNA photolyase that repairs UV-induced pyrimidine dimers with energy provided by visible light. We show that a Gc phrB mutant is not more sensitive to UV irradiation, independent of visible light exposure, and that the Gc phrB cannot complement an Escherichia coli phrB mutant strain. The Gc phrB mutant had a reduced colony size that was not a result of a growth defect and the mutant cells exhibited an altered morphology. Although the phrB mutant exhibited increased sensitivity to oxidative killing; it showed increased survival on media containing nalidixic acid or rifampicin, but did not have an increased mutation rate to these antibiotics or spectinomycin and kasugamycin. The Gc phrB mutant showed increased negative DNA supercoiling, but while the protein bound double-stranded DNA, it did not express topoisomerase activity. We conclude that the Gc PhrB has a previously unrecognized role in maintaining DNA supercoiling that is important for normal cell physiology.
Northwestern University, Feinberg School of Medicine, Department of Microbiology-Immunology, 303 E. Chicago Ave., Chicago, IL 60611, USA.
Laty A
LA
Cahoon
Elizabeth A
EA
Stohl
H Steven
HS
Seifert
D000595:Amino Acid Sequence; D001426:Bacterial Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D015169:Colony Count, Microbial; D004269:DNA, Bacterial / Q000378:metabolism; D004278:DNA, Superhelical / Q000378:metabolism*; D004255:Deoxyribodipyrimidine Photo-Lyase / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D004789:Enzyme Activation / Q000528:radiation effects; D004926:Escherichia coli / Q000528:radiation effects; D019143:Evolution, Molecular; D015964:Gene Expression Regulation, Bacterial / Q000187:drug effects / Q000528:radiation effects; D006801:Humans; D008027:Light; D050296:Microbial Viability / Q000528:radiation effects; D008969:Molecular Sequence Data; D009268:Nalidixic Acid / Q000494:pharmacology; D009344:Neisseria gonorrhoeae / Q000201:enzymology* / Q000235:genetics / Q000254:growth & development / Q000528:radiation effects; D009690:Nucleic Acid Conformation; D018384:Oxidative Stress / Q000187:drug effects / Q000528:radiation effects; D010641:Phenotype; D010802:Phylogeny; D017386:Sequence Homology, Amino Acid; D000198:Spectinomycin / Q000494:pharmacology
D001426:Bacterial Proteins; D004269:DNA, Bacterial; D004278:DNA, Superhelical; D009268:Nalidixic Acid; D000198:Spectinomycin; D004255:Deoxyribodipyrimidine Photo-Lyase
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