Compiling a molecular inventory for Mycobacterium bovis BCG at two growth rates: evidence for growth rate-mediated regulation of ribosome biosynthesis and lipid metabolism.
An experimental system of Mycobacterium tuberculosis growth in a carbon-limited chemostat has been established by the use of Mycobacterium bovis BCG as a model organism. For this model, carbon-limited chemostats with low concentrations of glycerol were used to simulate possible growth rates during different stages of tuberculosis. A doubling time of 23 h (D = 0.03 h(-1)) was adopted to represent cells during the acute phase of infection, whereas a lower dilution rate equivalent to a doubling time of 69 h (D = 0.01 h(-1)) was used to model mycobacterial persistence. This chemostat model allowed the specific response of the mycobacterial cell to carbon limitation at different growth rates to be elucidated. The macromolecular (RNA, DNA, carbohydrate, and lipid) and elemental (C, H, and N) compositions of the biomass were determined for steady-state cultures, revealing that carbohydrates and lipids comprised more than half of the dry mass of the BCG cell, with only a quarter of the dry weight consisting of protein and RNA. Consistent with studies of other bacteria, the specific growth rate impacts on the macromolecular content of BCG and the proportions of lipid, RNA, and protein increased significantly with the growth rate. The correlation of RNA content with the growth rate indicates that ribosome production in carbon-limited M. bovis BCG cells is subject to growth rate-dependent control. The results also clearly show that the proportion of lipids in the mycobacterial cell is very sensitive to changes in the growth rate, probably reflecting changes in the amounts of storage lipids. Finally, this study demonstrates the utility of the chemostat model of mycobacterial growth for functional genomic, physiology, and systems biology studies.
Beste
D J V
DJ
School of Biomedical and Molecular Sciences, University of Surrey, Guildford GU2 7XH, United Kingdom.
Peters
J
J
Hooper
T
T
Avignone-Rossa
C
C
Bushell
M E
ME
McFadden
J
J
D003470:Culture Media; D050356:Lipid Metabolism; D046911:Macromolecular Substances / Q000378:metabolism; D009163:Mycobacterium bovis / Q000254:growth & development* / Q000378:metabolism*; D012270:Ribosomes / Q000378:metabolism*
D003470:Culture Media; D046911:Macromolecular Substances
Tuberculosis (Edinb). 2004;84(3-4):228-38
15207492
J Theor Biol. 1982 Feb 7;94(3):651-70
6176814
Am J Clin Pathol. 1993 Jul;100(1):52-6
8346737
Biochem J. 1970 Apr;117(2):355-68
4911750
J Biol Stand. 1988 Oct;16(4):259-67
3058708
Microbiol Mol Biol Rev. 1998 Sep;62(3):646-66
9729604
Ir J Med Sci. 1971 Jan;140(1):16-25
4925435
J Exp Med. 2003 Sep 1;198(5):693-704
12953091
Clin Infect Dis. 1997 Jun;24(6):1139-46
9195072
Infect Immun. 1999 Jun;67(6):2867-73
10338493
J Gen Microbiol. 1979 Feb;110(2):431-41
220379
J Bacteriol. 1999 Jul;181(14):4326-33
10400591
J Biol Chem. 2003 Nov 21;278(47):46446-51
12963713
FEMS Microbiol Lett. 2000 Jul 15;188(2):141-6
10913697
Am J Dis Child. 1976 Apr;130(4):433-6
773173
Mol Microbiol. 2002 Feb;43(3):717-31
11929527
Biochim Biophys Acta. 1968 Nov 20;169(1):103-16
5727125
J Gen Microbiol. 1956 Aug;15(1):1-18
13357710
Infect Immun. 2002 Jan;70(1):286-91
11748194
J Bacteriol. 1956 Aug;72(2):132-41
13366889
Biochem J. 1967 Jun;103(3):67P
6049379
Curr Opin Microbiol. 2002 Jun;5(3):318-22
12057688
Proc Natl Acad Sci U S A. 2003 Jan 7;100(1):241-6
12506197
J Biotechnol. 2002 Jul 3;96(3):259-70
12044554
J Gen Microbiol. 1958 Dec;19(3):592-606
13611202
Biochem J. 1968 Nov;110(2):8P-9P
5726224
J Gen Microbiol. 1956 Jul;14(3):601-22
13346021
Arch Microbiol. 1986 Mar;144(2):105-9
3087325
J Bacteriol. 1999 Apr;181(7):2252-6
10094705
J Gen Microbiol. 1988 Aug;134(8):2111-21
3075652
Science. 1999 May 28;284(5419):1520-3
10348738
Scand J Infect Dis. 1997;29(5):526-7
9435049
Biotechnol Bioeng. 2001 Sep;76(2):132-43
11505383
Infect Immun. 1996 Jun;64(6):2062-9
8675308
J Mol Biol. 1966 Jul;18(2):308-20
5338757
J Appl Microbiol. 2000 Apr;88(4):669-77
10792526
J Bacteriol. 1974 Jul;119(1):270-81
4600702
Nature. 1951 Jul 28;168(4265):167
14875032
Electrophoresis. 2001 Aug;22(14):2936-46
11565788
Nature. 1998 Jun 11;393(6685):537-44
9634230
Am Rev Respir Dis. 1983 Feb;127(2):193-7
6131634
Nucleic Acids Res. 2003 Jan 1;31(1):187-9
12519978
Tuberculosis (Edinb). 2004;84(3-4):205-17
15207490
Microbiology (Reading). 2002 Oct;148(Pt 10):3129-3138
12368446
Microbiology (Reading). 2003 Mar;149(Pt 3):729-737
12634341
Microbiology (Reading). 1995 Dec;141 ( Pt 12):3105-11
8574403
Nature. 2000 Aug 17;406(6797):735-8
10963599
Appl Environ Microbiol. 2004 Apr;70(4):2354-66
15066832
Microbios. 1987;52(211):97-103
3695991
J Antimicrob Chemother. 2000 Dec;46(6):917-9
11102410
Infect Immun. 1991 Jun;59(6):1905-10
1903768
Lancet. 1980 Dec 20-27;2(8208-8209):1332-4
6109152
Arch Dis Child. 1988 Oct;63(10):1268-9
3196057
Indian J Chest Dis. 1974 Sep;16 Suppl:199-207
4442929
J Bacteriol. 1998 Nov;180(21):5756-61
9791129
Nat Med. 2002 Oct;8(10):1171-4
12219086
J Gen Microbiol. 1970 Sep;63(1):29-39
5500024
J Bacteriol. 1973 Jul;115(1):177-87
4577741
J Bacteriol. 1968 Feb;95(2):272-9
4966539
Microbiology (Reading). 2004 May;150(Pt 5):1413-1426
15133103
J Biol Chem. 1951 Nov;193(1):265-75
14907713