Mycobacterium tuberculosis WhiB4 regulates oxidative stress response to modulate survival and dissemination in vivo.
Host-generated oxidative stress is considered one of the main mechanisms constraining Mycobacterium tuberculosis (Mtb) growth. The redox-sensing mechanisms in Mtb are not completely understood. Here we show that WhiB4 responds to oxygen (O₂) and nitric oxide (NO) via its 4Fe-4S cluster and controls the oxidative stress response in Mtb. The WhiB4 mutant (MtbΔwhiB4) displayed an altered redox balance and a reduced membrane potential. Microarray analysis demonstrated that MtbΔwhiB4 overexpresses the antioxidant systems including alkyl hydroperoxidase (ahpC-ahpD) and rubredoxins (rubA-rubB). DNA binding assays showed that WhiB4 [4Fe-4S] cluster is dispensable for DNA binding. However, oxidation of the apo-WhiB4 Cys thiols induced disulphide-linked oligomerization, DNA binding and transcriptional repression, whereas reduction reversed the effect. Furthermore, WhiB4 binds DNA with a preference for GC-rich sequences. Expression analysis showed that oxidative stress repressed whiB4 and induced antioxidants in Mtb, while their hyper-induction was observed in MtbΔwhiB4. MtbΔwhiB4 showed increased resistance to oxidative stress in vitro and enhanced survival inside the macrophages. Lastly, MtbΔwhiB4 displayed hypervirulence in the lungs of guinea pigs, but showed a defect in dissemination to their spleen. These findings suggest that WhiB4 systematically calibrates the activation of oxidative stress response in Mtb to maintain redox balance, and to modulate virulence.
Immunology Group, International Centre for Genetic Engineering and Biotechnology, New Delhi 110067, India.
Manbeena
M
Chawla
Pankti
P
Parikh
Alka
A
Saxena
Mohamedhusen
M
Munshi
Mansi
M
Mehta
Deborah
D
Mai
Anup K
AK
Srivastava
K V
KV
Narasimhulu
Kevin E
KE
Redding
Nimi
N
Vashi
Dhiraj
D
Kumar
Adrie J C
AJ
Steyn
Amit
A
Singh
D000818:Animals; D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism*; D004268:DNA-Binding Proteins / Q000235:genetics / Q000378:metabolism; D017353:Gene Deletion; D020869:Gene Expression Profiling; D015964:Gene Expression Regulation, Bacterial; D006168:Guinea Pigs; D008168:Lung / Q000382:microbiology; D008264:Macrophages / Q000276:immunology / Q000382:microbiology; D046228:Microarray Analysis; D050296:Microbial Viability; D009169:Mycobacterium tuberculosis / Q000235:genetics* / Q000472:pathogenicity*; D009569:Nitric Oxide / Q000633:toxicity; D016877:Oxidants / Q000633:toxicity; D018384:Oxidative Stress; D010100:Oxygen / Q000633:toxicity; D013154:Spleen / Q000382:microbiology; D013312:Stress, Physiological; D014774:Virulence; D037521:Virulence Factors / Q000235:genetics / Q000378:metabolism
D001426:Bacterial Proteins; D004268:DNA-Binding Proteins; D016877:Oxidants; D037521:Virulence Factors; D009569:Nitric Oxide; D010100:Oxygen