A characterization of mutants of Staphylococcus aureus exhibiting increased sensitivity to ultraviolet radiation

Goering, Richard V

doi:10.31274/rtd-180813-3490

Cited by 2 publications

(4 citation statements)

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“…While ngr-374 behaves very similarly to the xthA mutations in E. coli, ngr-374, unlike xthA mutations, causes sensitivity to UV irradiation. This UV sensitivity implies that S. aureus has an AP endonuclease-dependent UV repair process which is distinct from the UV excision repair process detected by Goering and Pattee (11). ACKNOWLEDGMENTS This investigation was supported by National Science Foundation grants PCM-8110058 and PCM-8310681 and by Public Health Service grant AI-06647 from the National Institute of Allergy and Infectious Diseases.…”

Section: S Aureus Ap Endonuclease 713mentioning

confidence: 82%

“…NQO mimics the biological effect of UV irradiation so closely that mutants impaired in UV excision repair in E. coli and B. subtilis are sensitive to this compound (3,9,15,18). Similarly, uvs mutants of S. aureus, like their E. coli and B. subtilis counterparts, are impaired in UV excision repair (except uvs-J and uvs-8 mutants [11]) and are also sensitive to NQO (unpublished results). In contrast, ngr-374-carrying strains were not sensitive to NQO, indicating that ngr-374 was not related to the uvs mutations.…”

Section: S Aureus Ap Endonuclease 713mentioning

confidence: 85%

“…However, Wyman et al (48) reported that neither RN981 nor the Rec+ revertant of this mutant, RN1441, was impaired in its ability to reactivate UVirradiated bacteriophage; both strains carry ngr-374. The ngr-374-carrying strains failed to demonstrate any of the phenotypes characteristic of uvs mutations; therefore, it would appear that the UV sensitivity caused by ngr-374 may be due to an AP endonuclease-dependent UV repair process in S. aureus which is independent of the UV excision repair pathway affected by the uvs mutations described by Goering and Pattee (11).…”

Section: S Aureus Ap Endonuclease 713mentioning

confidence: 88%

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Characterization and genetic mapping of a mutation affecting apurinic endonuclease activity in Staphylococcus aureus

Tam¹,

Pattee²

1986

J Bacteriol

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Protoplast fusion between the Rec-mutant RN981 (L. Wyman, R. V. Goering, and R. P. Novick, Genetics 76:681-702, 1974) of Staphylococcus aureus NCTC 8325 and a Rec+ NCTC 8325 derivative yielded Rec+ recombinants that exhibited the increased sensitivity to N-methyl-N'-nitro-N-nitrosoguanidine characteristic of RN981. Transformation analyses identified a specific mutation, designated ngr-374, that was responsible not only for N-methyl-N'-nitro-N-nitrosoguanidine sensitivity, but also sensitivity to methyl methanesulfonate, ethyl methanesulfonate, nitrous acid, and UV irradiation. However, ngr-374-carrying recombinants shotved no significant increase in their sensitivity to mitomycin C or 4-nitroquinoline 1-oxide and were unaffected in recombination proficiency. In vitro assays showed that ngr-374-carrying strains had lower apurinic/apyrimidinic endonuclease activities than the wild type. The chromosomal locus occupied by ngr-374 was shown to exist in the gene order fl(Chr::TnSSl)40-ngr-374-thrBl06. In E. coli, three endonucleases with AP endonuclease activity have been characterized. Exonuclease III is responsible for 90% of the AP endonucleolytic activity in E. coli, and endonucleases III and IV account for the remainder (13,22,44). Generally, exonuclease III-deficient mutants, which carry the xthA mutation, are sensitive to methyl methanesulfonate (MMS) and nitrous acid, but are unchanged in their sensitivity to UV irradiation (4,8,23). Although these mutants lack the major AP endonuclease, they show no increase in their spontaneous mutation rate (49). Apparently, the minor AP endonucleases, endonucleases III and IV, supply sufficient AP endonucleolytic activity to repair the spontaneously occurring AP sites; however, they are unable to manage the increased number of AP sites formed with MMS or nitrous acid treatments (22,44,46).In this report, we describe ngr-374, a mutation impairing AP endonuclease activity and originating in the recombination-deficient (Rec-) mutant RN981 (48) acid. ngr-374-carrying strains were also sensitive to UV irradiation, suggesting that S. aureus may have an AP endonuclease-dependent DNA repair mechanism for UVgenerated DNA lesions. MATERIALS AND METHODSBacterial strains. The strains of S. aureus, Bacillus subtilis, and E. coli used in this study are described in Table 1. All cultures were maintained as described previously (35).Culture media. All commercially available dehydrated culture media were supplemented with thymine (20 ,ug/ml) and adenine, guanine, cytosine, and uracil (5 ,ug/ml each). The composition of complete defined synthetic agar (31) was modified by omitting the appropriate amino acids, purines, and pyrimidines as needed to score and select nutritional markers (2,29,30). N-Methyl-N'-nitro-N-nitrosoguanidine (MNNG) agar was prepared by adding 5 ,ug of MNNG (Sigma Chemical Co.), freshly dissolved in absolute methanol, per ml to autoclaved, cooled (45°C) Trypticase soy agar (TSA; BBL Microbiology Systems) rehydrated in 100 mM KH2PO4, pH 6.0 (10). MNNG agar was used on...

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Section: S Aureus Ap Endonuclease 713mentioning

confidence: 82%

Section: S Aureus Ap Endonuclease 713mentioning

confidence: 85%

Section: S Aureus Ap Endonuclease 713mentioning

confidence: 88%

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Characterization and genetic mapping of a mutation affecting apurinic endonuclease activity in Staphylococcus aureus

Tam¹,

Pattee²

1986

J Bacteriol

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“…Alkylation leading to point mutations (5,8) extremely laborious and expensive. However, with current sequencing technology, whole bacterial genomes are now sequenced in days at very affordable prices.…”

Section: Introductionmentioning

confidence: 99%

Chemical and UV Mutagenesis

Bose

2014

Methods in Molecular Biology

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The ability to create mutations is an important step towards understanding bacterial physiology and virulence. While targeted approaches are invaluable, the ability to produce genome-wide random mutations can lead to crucial discoveries. Transposon mutagenesis is a useful approach, but many interesting mutations can be missed by these insertions that interrupt coding and noncoding sequences due to the integration of an entire transposon. Chemical mutagenesis and UV-based random mutagenesis are alternate approaches to isolate mutations of interest with the potential of only single nucleotide changes. Once a standard method, difficulty in identifying mutation sites had decreased the popularity of this technique. However, thanks to the recent emergence of economical whole-genome sequencing, this approach to making mutations can once again become a viable option. Therefore, this chapter provides an overview protocol for random mutagenesis using UV light or DNA-damaging chemicals.

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A characterization of mutants of Staphylococcus aureus exhibiting increased sensitivity to ultraviolet radiation

Cited by 2 publications

References 49 publications

Characterization and genetic mapping of a mutation affecting apurinic endonuclease activity in Staphylococcus aureus

Characterization and genetic mapping of a mutation affecting apurinic endonuclease activity in Staphylococcus aureus

Chemical and UV Mutagenesis

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