Identification of a chemical that inhibits the mycobacterial UvrABC complex in nucleotide excision repair

Nayef Mazloum, Melanie A. Stegman, Deborah L. Croteau, Bennett Van Houten, Nyoun Soo Kwon, Yan Ling, Caitlyn Dickinson, Aditya Venugopal, Mohammad Atif Towheed, Carl Nathan

Research output: Contribution to journalArticle

16 Citations (Scopus)

Abstract

Bacterial DNA can be damaged by reactive nitrogen and oxygen intermediates (RNI and ROI) generated by host immunity, as well as by antibiotics that trigger bacterial production of ROI. Thus a pathogens ability to repair its DNA may be important for persistent infection. A prominent role for nucleotide excision repair (NER) in disease caused by Mycobacterium tuberculosis (Mtb) was suggested by attenuation of uvrB-deficient Mtb in mice. However, it was unknown if Mtbs Uvr proteins could execute NER. Here we report that recombinant UvrA, UvrB, and UvrC from Mtb collectively bound and cleaved plasmid DNA exposed to ultraviolet (UV) irradiation or peroxynitrite. We used the DNA incision assay to test the mechanism of action of compounds identified in a high-throughput screen for their ability to delay recovery of M. smegmatis from UV irradiation. 2-(5-Amino-1,3,4-thiadiazol-2-ylbenzo[f]chromen-3-one) (ATBC) but not several closely related compounds inhibited cleavage of damaged DNA by UvrA, UvrB, and UvrC without intercalating in DNA and impaired recovery of M. smegmatis from UV irradiation. ATBC did not affect bacterial growth in the absence of UV exposure, nor did it exacerbate the growth defect of UV-irradiated mycobacteria that lacked uvrB. Thus, ATBC appears to be a cell-penetrant, selective inhibitor of mycobacterial NER. Chemical inhibitors of NER may facilitate studies of the role of NER in prokaryotic pathobiology.

Original languageEnglish
Pages (from-to)1329-1335
Number of pages7
JournalBiochemistry
Volume50
Issue number8
DOIs
Publication statusPublished - 1 Mar 2011
Externally publishedYes

Fingerprint

DNA Repair
Repair
Nucleotides
Smegma
Mycobacterium tuberculosis
DNA
Irradiation
Recovery
Bacterial DNA
DNA Cleavage
Peroxynitrous Acid
Pathogens
Mycobacterium
Growth
Immunity
Assays
Plasmids
Nitrogen
Throughput
Oxygen

ASJC Scopus subject areas

  • Biochemistry

Cite this

Mazloum, N., Stegman, M. A., Croteau, D. L., Van Houten, B., Kwon, N. S., Ling, Y., ... Nathan, C. (2011). Identification of a chemical that inhibits the mycobacterial UvrABC complex in nucleotide excision repair. Biochemistry, 50(8), 1329-1335. https://doi.org/10.1021/bi101674c

Identification of a chemical that inhibits the mycobacterial UvrABC complex in nucleotide excision repair. / Mazloum, Nayef; Stegman, Melanie A.; Croteau, Deborah L.; Van Houten, Bennett; Kwon, Nyoun Soo; Ling, Yan; Dickinson, Caitlyn; Venugopal, Aditya; Towheed, Mohammad Atif; Nathan, Carl.

In: Biochemistry, Vol. 50, No. 8, 01.03.2011, p. 1329-1335.

Research output: Contribution to journalArticle

Mazloum, N, Stegman, MA, Croteau, DL, Van Houten, B, Kwon, NS, Ling, Y, Dickinson, C, Venugopal, A, Towheed, MA & Nathan, C 2011, 'Identification of a chemical that inhibits the mycobacterial UvrABC complex in nucleotide excision repair', Biochemistry, vol. 50, no. 8, pp. 1329-1335. https://doi.org/10.1021/bi101674c
Mazloum, Nayef ; Stegman, Melanie A. ; Croteau, Deborah L. ; Van Houten, Bennett ; Kwon, Nyoun Soo ; Ling, Yan ; Dickinson, Caitlyn ; Venugopal, Aditya ; Towheed, Mohammad Atif ; Nathan, Carl. / Identification of a chemical that inhibits the mycobacterial UvrABC complex in nucleotide excision repair. In: Biochemistry. 2011 ; Vol. 50, No. 8. pp. 1329-1335.
@article{9349cfe703f740aa80e7d507d9745ffe,
title = "Identification of a chemical that inhibits the mycobacterial UvrABC complex in nucleotide excision repair",
abstract = "Bacterial DNA can be damaged by reactive nitrogen and oxygen intermediates (RNI and ROI) generated by host immunity, as well as by antibiotics that trigger bacterial production of ROI. Thus a pathogens ability to repair its DNA may be important for persistent infection. A prominent role for nucleotide excision repair (NER) in disease caused by Mycobacterium tuberculosis (Mtb) was suggested by attenuation of uvrB-deficient Mtb in mice. However, it was unknown if Mtbs Uvr proteins could execute NER. Here we report that recombinant UvrA, UvrB, and UvrC from Mtb collectively bound and cleaved plasmid DNA exposed to ultraviolet (UV) irradiation or peroxynitrite. We used the DNA incision assay to test the mechanism of action of compounds identified in a high-throughput screen for their ability to delay recovery of M. smegmatis from UV irradiation. 2-(5-Amino-1,3,4-thiadiazol-2-ylbenzo[f]chromen-3-one) (ATBC) but not several closely related compounds inhibited cleavage of damaged DNA by UvrA, UvrB, and UvrC without intercalating in DNA and impaired recovery of M. smegmatis from UV irradiation. ATBC did not affect bacterial growth in the absence of UV exposure, nor did it exacerbate the growth defect of UV-irradiated mycobacteria that lacked uvrB. Thus, ATBC appears to be a cell-penetrant, selective inhibitor of mycobacterial NER. Chemical inhibitors of NER may facilitate studies of the role of NER in prokaryotic pathobiology.",
author = "Nayef Mazloum and Stegman, {Melanie A.} and Croteau, {Deborah L.} and {Van Houten}, Bennett and Kwon, {Nyoun Soo} and Yan Ling and Caitlyn Dickinson and Aditya Venugopal and Towheed, {Mohammad Atif} and Carl Nathan",
year = "2011",
month = "3",
day = "1",
doi = "10.1021/bi101674c",
language = "English",
volume = "50",
pages = "1329--1335",
journal = "Biochemistry",
issn = "0006-2960",
publisher = "American Chemical Society",
number = "8",

}

TY - JOUR

T1 - Identification of a chemical that inhibits the mycobacterial UvrABC complex in nucleotide excision repair

AU - Mazloum, Nayef

AU - Stegman, Melanie A.

AU - Croteau, Deborah L.

AU - Van Houten, Bennett

AU - Kwon, Nyoun Soo

AU - Ling, Yan

AU - Dickinson, Caitlyn

AU - Venugopal, Aditya

AU - Towheed, Mohammad Atif

AU - Nathan, Carl

PY - 2011/3/1

Y1 - 2011/3/1

N2 - Bacterial DNA can be damaged by reactive nitrogen and oxygen intermediates (RNI and ROI) generated by host immunity, as well as by antibiotics that trigger bacterial production of ROI. Thus a pathogens ability to repair its DNA may be important for persistent infection. A prominent role for nucleotide excision repair (NER) in disease caused by Mycobacterium tuberculosis (Mtb) was suggested by attenuation of uvrB-deficient Mtb in mice. However, it was unknown if Mtbs Uvr proteins could execute NER. Here we report that recombinant UvrA, UvrB, and UvrC from Mtb collectively bound and cleaved plasmid DNA exposed to ultraviolet (UV) irradiation or peroxynitrite. We used the DNA incision assay to test the mechanism of action of compounds identified in a high-throughput screen for their ability to delay recovery of M. smegmatis from UV irradiation. 2-(5-Amino-1,3,4-thiadiazol-2-ylbenzo[f]chromen-3-one) (ATBC) but not several closely related compounds inhibited cleavage of damaged DNA by UvrA, UvrB, and UvrC without intercalating in DNA and impaired recovery of M. smegmatis from UV irradiation. ATBC did not affect bacterial growth in the absence of UV exposure, nor did it exacerbate the growth defect of UV-irradiated mycobacteria that lacked uvrB. Thus, ATBC appears to be a cell-penetrant, selective inhibitor of mycobacterial NER. Chemical inhibitors of NER may facilitate studies of the role of NER in prokaryotic pathobiology.

AB - Bacterial DNA can be damaged by reactive nitrogen and oxygen intermediates (RNI and ROI) generated by host immunity, as well as by antibiotics that trigger bacterial production of ROI. Thus a pathogens ability to repair its DNA may be important for persistent infection. A prominent role for nucleotide excision repair (NER) in disease caused by Mycobacterium tuberculosis (Mtb) was suggested by attenuation of uvrB-deficient Mtb in mice. However, it was unknown if Mtbs Uvr proteins could execute NER. Here we report that recombinant UvrA, UvrB, and UvrC from Mtb collectively bound and cleaved plasmid DNA exposed to ultraviolet (UV) irradiation or peroxynitrite. We used the DNA incision assay to test the mechanism of action of compounds identified in a high-throughput screen for their ability to delay recovery of M. smegmatis from UV irradiation. 2-(5-Amino-1,3,4-thiadiazol-2-ylbenzo[f]chromen-3-one) (ATBC) but not several closely related compounds inhibited cleavage of damaged DNA by UvrA, UvrB, and UvrC without intercalating in DNA and impaired recovery of M. smegmatis from UV irradiation. ATBC did not affect bacterial growth in the absence of UV exposure, nor did it exacerbate the growth defect of UV-irradiated mycobacteria that lacked uvrB. Thus, ATBC appears to be a cell-penetrant, selective inhibitor of mycobacterial NER. Chemical inhibitors of NER may facilitate studies of the role of NER in prokaryotic pathobiology.

UR - http://www.scopus.com/inward/record.url?scp=79951862276&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=79951862276&partnerID=8YFLogxK

U2 - 10.1021/bi101674c

DO - 10.1021/bi101674c

M3 - Article

C2 - 21235228

AN - SCOPUS:79951862276

VL - 50

SP - 1329

EP - 1335

JO - Biochemistry

JF - Biochemistry

SN - 0006-2960

IS - 8

ER -