27. Carroll MW, Jeon D, Mountz JM, Lee JD,
Jeong YJ, Zia N, Lee M, Lee J, Via LE, Lee S,
Eum SY, Lee SJ, Goldfeder LC, Cai Y, Jin B,
Kim Y, Oh T, Chen RY, Dodd LE, Gu W,
Dartois V, Park SK, Kim CT, Barry CE 3rd,
Cho SN (2013) Efficacy and safety of metronidazole for pulmonary multidrug-resistant
tuberculosis. Antimicrob Agents Chemother
57(8):3903–3909. https://doi.org/10.1128/
AAC.00753-13
28. Coates A, Hu Y, Bax R, Page C (2002) The
future challenges facing the development of
new antimicrobial drugs. Nat Rev Drug Discov
1(11):895–910. https://doi.org/10.1038/
nrd940
29. Rao SP, Alonso S, Rand L, Dick T, Pethe K
(2008) The protonmotive force is required for
maintaining ATP homeostasis and viability of
hypoxic, nonreplicating Mycobacterium tuberculosis. Proc Natl Acad Sci U S A 105
(33):11945–11950.
https://doi.org/10.
1073/pnas.0711697105
30. Cho SH, Warit S, Wan B, Hwang CH, Pauli
GF, Franzblau SG (2007) Low-oxygen-recovery assay for high-throughput screening of
compounds against nonreplicating Mycobacterium tuberculosis. Antimicrob Agents Chemother 51(4):1380–1385. https://doi.org/
10.1128/AAC.00055-06
31. Gold B, Roberts J, Ling Y, Quezada LL,
Glasheen J, Ballinger E, Somersan-Karakaya S,
Warrier T, Warren JD, Nathan C (2015) Rapid,
semiquantitative assay to discriminate among
compounds with activity against replicating or
nonreplicating Mycobacterium tuberculosis.
Antimicrob
Agents
Chemother
59
(10):6521–6538. https://doi.org/10.1128/
AAC.00803-15
32. Gold B, Roberts J, Ling Y, Lopez Quezada L,
Glasheen J, Ballinger E, Somersan-Karakaya S,
Warrier T, Nathan C (2016) Visualization of
the charcoal agar resazurin assay for semiquantitative, medium-throughput enumeration of mycobacteria. J Vis Exp (118).
https://doi.org/10.3791/54690
33. MacMicking JD, Taylor GA, McKinney JD
(2003) Immune control of tuberculosis by
IFN-gamma-inducible LRG-47. Science 302
(5645):654–659
34. MacMicking JD, North RJ, LaCourse R, Mudgett JS, Shah SK, Nathan CF (1997) Identification of nitric oxide synthase as a protective
locus against tuberculosis. Proc Natl Acad Sci
U S A 94(10):5243–5248
35. Cunningham-Bussel A, Zhang T, Nathan CF
(2013) Nitrite produced by Mycobacterium
tuberculosis in human macrophages in physiologic oxygen impacts bacterial
ATP
consumption and gene expression. Proc Natl
Acad Sci U S A 110(45):E4256–E4265.
https://doi.org/10.1073/pnas.1316894110
36. Aly S, Wagner K, Keller C, Malm S, Malzan A,
Brandau S, Bange FC, Ehlers S (2006) Oxygen
status of lung granulomas in Mycobacterium
tuberculosis-infected mice. J Pathol 210
(3):298–305. https://doi.org/10.1002/path.
2055
37. Marrero J, Rhee KY, Schnappinger D, Pethe K,
Ehrt S (2010) Gluconeogenic carbon flow of
tricarboxylic acid cycle intermediates is critical
for Mycobacterium tuberculosis to establish and
maintain infection. Proc Natl Acad Sci U S A
107(21):9819–9824.
https://doi.org/10.
1073/pnas.1000715107
38. Munoz-Elias EJ, McKinney JD (2005) Mycobacterium tuberculosis isocitrate lyases 1 and
2 are jointly required for in vivo growth and
virulence. Nat Med 11(6):638–644. https://
doi.org/10.1038/nm1252
39. Gold B, Rodriguez GM, Marras SA,
Pentecost M, Smith I (2001) The Mycobacterium tuberculosis IdeR is a dual functional regulator that controls transcription of genes
involved in iron acquisition, iron storage and
survival in macrophages. Mol Microbiol 42
(3):851–865.
https://doi.org/10.1046/j.
1365-2958.2001.02684.x
40. Rodriguez GM, Voskuil MI, Gold B, Schoolnik
GK, Smith I (2002) ideR, an essential gene in
Mycobacterium tuberculosis: role of IdeR in
iron-dependent gene expression, iron metabolism, and oxidative stress response. Infect
Immun 70(7):3371–3381. https://doi.org/
10.1128/iai.70.7.3371-3381.2002
41. Timm J, Post FA, Bekker LG, Walther GB,
Wainwright HC, Manganelli R, Chan WT,
Tsenova L, Gold B, Smith I, Kaplan G, McKinney JD (2003) Differential expression of iron-,
carbon-, and oxygen-responsive mycobacterial
genes in the lungs of chronically infected mice
and tuberculosis patients. Proc Natl Acad Sci U
S A 100(24):14321–14326. https://doi.org/
10.1073/pnas.2436197100
42. Walters SB, Dubnau E, Kolesnikova I, Laval F,
Daffe M, Smith I (2006) The Mycobacterium
tuberculosis PhoPR two-component system
regulates genes essential for virulence and complex lipid biosynthesis. Mol Microbiol 60
(2):312–330.
https://doi.org/10.1111/j.
1365-2958.2006.05102.x
43. Hondalus MK, Bardarov S, Russell R, Chan J,
Jacobs WR Jr, Bloom BR (2000) Attenuation
of and protection induced by a leucine auxotroph of Mycobacterium tuberculosis. Infect
Immun 68(5):2888–2898
634
Ben Gold et al.
Jeong YJ, Zia N, Lee M, Lee J, Via LE, Lee S,
Eum SY, Lee SJ, Goldfeder LC, Cai Y, Jin B,
Kim Y, Oh T, Chen RY, Dodd LE, Gu W,
Dartois V, Park SK, Kim CT, Barry CE 3rd,
Cho SN (2013) Efficacy and safety of metronidazole for pulmonary multidrug-resistant
tuberculosis. Antimicrob Agents Chemother
57(8):3903–3909. https://doi.org/10.1128/
AAC.00753-13
28. Coates A, Hu Y, Bax R, Page C (2002) The
future challenges facing the development of
new antimicrobial drugs. Nat Rev Drug Discov
1(11):895–910. https://doi.org/10.1038/
nrd940
29. Rao SP, Alonso S, Rand L, Dick T, Pethe K
(2008) The protonmotive force is required for
maintaining ATP homeostasis and viability of
hypoxic, nonreplicating Mycobacterium tuberculosis. Proc Natl Acad Sci U S A 105
(33):11945–11950.
https://doi.org/10.
1073/pnas.0711697105
30. Cho SH, Warit S, Wan B, Hwang CH, Pauli
GF, Franzblau SG (2007) Low-oxygen-recovery assay for high-throughput screening of
compounds against nonreplicating Mycobacterium tuberculosis. Antimicrob Agents Chemother 51(4):1380–1385. https://doi.org/
10.1128/AAC.00055-06
31. Gold B, Roberts J, Ling Y, Quezada LL,
Glasheen J, Ballinger E, Somersan-Karakaya S,
Warrier T, Warren JD, Nathan C (2015) Rapid,
semiquantitative assay to discriminate among
compounds with activity against replicating or
nonreplicating Mycobacterium tuberculosis.
Antimicrob
Agents
Chemother
59
(10):6521–6538. https://doi.org/10.1128/
AAC.00803-15
32. Gold B, Roberts J, Ling Y, Lopez Quezada L,
Glasheen J, Ballinger E, Somersan-Karakaya S,
Warrier T, Nathan C (2016) Visualization of
the charcoal agar resazurin assay for semiquantitative, medium-throughput enumeration of mycobacteria. J Vis Exp (118).
https://doi.org/10.3791/54690
33. MacMicking JD, Taylor GA, McKinney JD
(2003) Immune control of tuberculosis by
IFN-gamma-inducible LRG-47. Science 302
(5645):654–659
34. MacMicking JD, North RJ, LaCourse R, Mudgett JS, Shah SK, Nathan CF (1997) Identification of nitric oxide synthase as a protective
locus against tuberculosis. Proc Natl Acad Sci
U S A 94(10):5243–5248
35. Cunningham-Bussel A, Zhang T, Nathan CF
(2013) Nitrite produced by Mycobacterium
tuberculosis in human macrophages in physiologic oxygen impacts bacterial
ATP
consumption and gene expression. Proc Natl
Acad Sci U S A 110(45):E4256–E4265.
https://doi.org/10.1073/pnas.1316894110
36. Aly S, Wagner K, Keller C, Malm S, Malzan A,
Brandau S, Bange FC, Ehlers S (2006) Oxygen
status of lung granulomas in Mycobacterium
tuberculosis-infected mice. J Pathol 210
(3):298–305. https://doi.org/10.1002/path.
2055
37. Marrero J, Rhee KY, Schnappinger D, Pethe K,
Ehrt S (2010) Gluconeogenic carbon flow of
tricarboxylic acid cycle intermediates is critical
for Mycobacterium tuberculosis to establish and
maintain infection. Proc Natl Acad Sci U S A
107(21):9819–9824.
https://doi.org/10.
1073/pnas.1000715107
38. Munoz-Elias EJ, McKinney JD (2005) Mycobacterium tuberculosis isocitrate lyases 1 and
2 are jointly required for in vivo growth and
virulence. Nat Med 11(6):638–644. https://
doi.org/10.1038/nm1252
39. Gold B, Rodriguez GM, Marras SA,
Pentecost M, Smith I (2001) The Mycobacterium tuberculosis IdeR is a dual functional regulator that controls transcription of genes
involved in iron acquisition, iron storage and
survival in macrophages. Mol Microbiol 42
(3):851–865.
https://doi.org/10.1046/j.
1365-2958.2001.02684.x
40. Rodriguez GM, Voskuil MI, Gold B, Schoolnik
GK, Smith I (2002) ideR, an essential gene in
Mycobacterium tuberculosis: role of IdeR in
iron-dependent gene expression, iron metabolism, and oxidative stress response. Infect
Immun 70(7):3371–3381. https://doi.org/
10.1128/iai.70.7.3371-3381.2002
41. Timm J, Post FA, Bekker LG, Walther GB,
Wainwright HC, Manganelli R, Chan WT,
Tsenova L, Gold B, Smith I, Kaplan G, McKinney JD (2003) Differential expression of iron-,
carbon-, and oxygen-responsive mycobacterial
genes in the lungs of chronically infected mice
and tuberculosis patients. Proc Natl Acad Sci U
S A 100(24):14321–14326. https://doi.org/
10.1073/pnas.2436197100
42. Walters SB, Dubnau E, Kolesnikova I, Laval F,
Daffe M, Smith I (2006) The Mycobacterium
tuberculosis PhoPR two-component system
regulates genes essential for virulence and complex lipid biosynthesis. Mol Microbiol 60
(2):312–330.
https://doi.org/10.1111/j.
1365-2958.2006.05102.x
43. Hondalus MK, Bardarov S, Russell R, Chan J,
Jacobs WR Jr, Bloom BR (2000) Attenuation
of and protection induced by a leucine auxotroph of Mycobacterium tuberculosis. Infect
Immun 68(5):2888–2898
634
Ben Gold et al.