Identification of aminopyrimidine-sulfonamides as potent modulators of Wag31-mediated cell elongation in mycobacteria.
There is an urgent need to discover new anti-tubercular agents with novel mechanisms of action in order to tackle the scourge of drug-resistant tuberculosis. Here, we report the identification of such a molecule - an AminoPYrimidine-Sulfonamide (APYS1) that has potent, bactericidal activity against M. tuberculosis. Mutations in APYS1-resistant M. tuberculosis mapped exclusively to wag31, a gene that encodes a scaffolding protein thought to orchestrate cell elongation. Recombineering confirmed that a Gln201Arg mutation in Wag31 was sufficient to cause resistance to APYS1, however, neither overexpression nor conditional depletion of wag31 impacted M. tuberculosis susceptibility to this compound. In contrast, expression of the wildtype allele of wag31 in APYS1-resistant M. tuberculosis was dominant and restored susceptibility to APYS1 to wildtype levels. Time-lapse imaging and scanning electron microscopy revealed that APYS1 caused gross malformation of the old pole of M. tuberculosis, with eventual lysis. These effects resembled the morphological changes observed following transcriptional silencing of wag31 in M. tuberculosis. These data show that Wag31 is likely not the direct target of APYS1, but the striking phenotypic similarity between APYS1 exposure and genetic depletion of Wag31 in M. tuberculosis suggests that APYS1 might indirectly affect Wag31 through an as yet unknown mechanism.
Institute of Infectious Disease and Molecular Medicine & Department of Pathology, University of Cape Town, MRC/NHLS/UCT Molecular Mycobacteriology Research Unit & DST/NRF Centre of Excellence for Biomedical TB Research, South Africa.
Vinayak
V
Singh
Microbiology and Microsystems, Global Health Institute, Ecole Polytechnique Federale de Lausanne, Lausanne, Switzerland.
Neeraj
N
Dhar
Vichem Chemie Research Ltd, Herman, Otto u. 15, Budapest, 1022, Hungary.
János
J
Pató
Microbial Pathogenesis, Global Health Institute, Ecole Polytechnique Fédérale de Lausanne, Lausanne, Switzerland.
Gaëlle S
GS
Kolly
Now at: 1st Department of Internal Medicine, University of Cologne, Cologne, Germany.
Jana
J
Korduláková
Now at: 1st Department of Internal Medicine, University of Cologne, Cologne, Germany.
Martin
M
Forbak
Institute of Infectious Disease and Molecular Medicine & Department of Pathology, University of Cape Town, MRC/NHLS/UCT Molecular Mycobacteriology Research Unit & DST/NRF Centre of Excellence for Biomedical TB Research, South Africa.
Joanna C
JC
Evans
Microbial Pathogenesis, Global Health Institute, Ecole Polytechnique Fédérale de Lausanne, Lausanne, Switzerland.
Rita
R
Székely
Microbial Pathogenesis, Global Health Institute, Ecole Polytechnique Fédérale de Lausanne, Lausanne, Switzerland.|Now at: 1st Department of Internal Medicine, University of Cologne, Cologne, Germany.
Jan
J
Rybniker
Faculty of Natural Sciences, Department of Biochemistry, Comenius University in Bratislava, Bratislava, Slovakia.
Zuzana
Z
Palčeková
Faculty of Natural Sciences, Department of Biochemistry, Comenius University in Bratislava, Bratislava, Slovakia.
Júlia
J
Zemanová
Microbiology and Microsystems, Global Health Institute, Ecole Polytechnique Federale de Lausanne, Lausanne, Switzerland.
Isabella
I
Santi
Microbiology and Microsystems, Global Health Institute, Ecole Polytechnique Federale de Lausanne, Lausanne, Switzerland.
François
F
Signorino-Gelo
Departamento de Microbiología, Facultad de Medicina, Universidad de Zaragoza, and Fundación ARAID, Zaragoza, Spain; CIBERES, Instituto de Salud Carlos III, Madrid, Zaragoza, Spain.
Liliana
L
Rodrigues
Microbial Pathogenesis, Global Health Institute, Ecole Polytechnique Fédérale de Lausanne, Lausanne, Switzerland.
Anthony
A
Vocat
Department of Cell and Molecular Biology, Uppsala University, Biomedical Center, Uppsala, Sweden.
Adrian S
AS
Covarrubias
Institute of Chemical Sciences and Engineering (ISIC), Ecole Polytechnique Fédérale de Lausanne, Lausanne, Switzerland.
Monica G
MG
Rengifo
Institute of Chemical Sciences and Engineering (ISIC), Ecole Polytechnique Fédérale de Lausanne, Lausanne, Switzerland.
Kai
K
Johnsson
Department of Cell and Molecular Biology, Uppsala University, Biomedical Center, Uppsala, Sweden.
Sherry
S
Mowbray
Alere (San Diego), Summer Ridge Road, San Diego, CA, 92121, USA.
Joseph
J
Buechler
Center for Infection and Immunity, Inserm U1019, CNRS UMR8204, Institut Pasteur de Lille, Université de Lille, Lille, France.
Vincent
V
Delorme
Center for Infection and Immunity, Inserm U1019, CNRS UMR8204, Institut Pasteur de Lille, Université de Lille, Lille, France.
Priscille
P
Brodin
Interdisciplinary Centre for Electron Microscopy, Ecole Polytechnique Fédérale de Lausanne, Lausanne, Switzerland.
Graham W
GW
Knott
Departamento de Microbiología, Facultad de Medicina, Universidad de Zaragoza, and Fundación ARAID, Zaragoza, Spain; CIBERES, Instituto de Salud Carlos III, Madrid, Zaragoza, Spain.
José A
JA
Aínsa
Institute of Infectious Disease and Molecular Medicine & Department of Pathology, University of Cape Town, MRC/NHLS/UCT Molecular Mycobacteriology Research Unit & DST/NRF Centre of Excellence for Biomedical TB Research, South Africa.
Digby F
DF
Warner
Vichem Chemie Research Ltd, Herman, Otto u. 15, Budapest, 1022, Hungary.
György
G
Kéri
Faculty of Natural Sciences, Department of Biochemistry, Comenius University in Bratislava, Bratislava, Slovakia.
Katarína
K
Mikušová
Microbiology and Microsystems, Global Health Institute, Ecole Polytechnique Federale de Lausanne, Lausanne, Switzerland.
John D
JD
McKinney
Microbial Pathogenesis, Global Health Institute, Ecole Polytechnique Fédérale de Lausanne, Lausanne, Switzerland.
Stewart T
ST
Cole
Institute of Infectious Disease and Molecular Medicine & Department of Pathology, University of Cape Town, MRC/NHLS/UCT Molecular Mycobacteriology Research Unit & DST/NRF Centre of Excellence for Biomedical TB Research, South Africa.
Valerie
V
Mizrahi
Microbial Pathogenesis, Global Health Institute, Ecole Polytechnique Fédérale de Lausanne, Lausanne, Switzerland.
Ruben C
RC
Hartkoorn
D000900:Anti-Bacterial Agents / Q000493:pharmacokinetics; D000995:Antitubercular Agents / Q000378:metabolism*; D001426:Bacterial Proteins / Q000235:genetics* / Q000378:metabolism*; D049108:Cell Enlargement; D055808:Drug Discovery / Q000379:methods; D015964:Gene Expression Regulation, Bacterial / Q000235:genetics; D020102:Mycobacterium smegmatis / Q000235:genetics / Q000378:metabolism; D009169:Mycobacterium tuberculosis / Q000235:genetics / Q000378:metabolism; D011743:Pyrimidines / Q000737:chemistry / Q000378:metabolism / Q000493:pharmacokinetics*; D017386:Sequence Homology, Amino Acid; D013449:Sulfonamides / Q000378:metabolism / Q000493:pharmacokinetics; D059008:Time-Lapse Imaging
D000900:Anti-Bacterial Agents; D000995:Antitubercular Agents; D001426:Bacterial Proteins; D011743:Pyrimidines; D013449:Sulfonamides; C000591547:Wag31 protein, Mycobacterium tuberculosis; C012180:2-aminopyrimidine