SpoT-triggered stringent response controls usp gene expression in Pseudomonas aeruginosa.
The universal stress proteins (Usps) UspK (PA3309) and UspN (PA4352) of Pseudomonas aeruginosa are essential for surviving specific anaerobic energy stress conditions such as pyruvate fermentation and anaerobic stationary phase. Expression of the respective genes is under the control of the oxygen-sensing regulator Anr. In this study we investigated the regulation of uspN and three additional P. aeruginosa usp genes: uspL (PA1789), uspM (PA4328), and uspO (PA5027). Anr induces expression of these genes in response to anaerobic conditions. Using promoter-lacZ fusions, we showed that PuspL-lacZ, PuspM-lacZ, and PuspO-lacZ were also induced in stationary phase as described for PuspN-lacZ. However, stationary phase gene expression was abolished in the P. aeruginosa triple mutant Deltaanr DeltarelA DeltaspoT. The relA and spoT genes encode the regulatory components of the stringent response. We determined pppGpp and ppGpp levels using a thin-layer chromatography approach and detected the accumulation of ppGpp in the wild type and the DeltarelA mutant in stationary phase, indicating a SpoT-derived control of ppGpp accumulation. Additional investigation of stationary phase in LB medium revealed that alkaline pH values are involved in the regulatory process of ppGpp accumulation.
Boes
Nelli
N
Institute of Microbiology, Technische Universität Braunschweig, Spielmannstr. 7, D-38106 Braunschweig, Germany.
Schreiber
Kerstin
K
Schobert
Max
M
D000693:Anaerobiosis; D001426:Bacterial Proteins / Q000235:genetics* / Q000378:metabolism / Q000502:physiology; D002855:Chromatography, Thin Layer; D015964:Gene Expression Regulation, Bacterial; D006158:Guanosine Pentaphosphate / Q000378:metabolism; D006159:Guanosine Tetraphosphate / Q000378:metabolism; D006863:Hydrogen-Ion Concentration; D008954:Models, Biological; D011401:Promoter Regions, Genetic / Q000235:genetics; D011550:Pseudomonas aeruginosa / Q000235:genetics* / Q000378:metabolism; D015534:Trans-Activators / Q000235:genetics / Q000378:metabolism / Q000502:physiology; D001616:beta-Galactosidase / Q000235:genetics / Q000378:metabolism
D001426:Bacterial Proteins; D015534:Trans-Activators; C072908:ANR protein, Pseudomonas aeruginosa; D006159:Guanosine Tetraphosphate; D006158:Guanosine Pentaphosphate; D001616:beta-Galactosidase
Mol Microbiol. 2007 Jul;65(1):153-65
17581126
Genes Dev. 2002 May 15;16(10):1260-70
12023304
Proc Natl Acad Sci U S A. 2006 Feb 21;103(8):2833-8
16477005
Microbiology (Reading). 2002 Feb;148(Pt 2):559-570
11832519
Mol Microbiol. 2007 Aug;65(4):968-78
17640273
Mol Gen Genet. 1980;178(2):325-7
6156378
Dev Cell. 2002 Oct;3(4):593-603
12408810
BMC Genomics. 2006 Sep 08;7:230
16961923
Bioinformatics. 2005 Nov 15;21(22):4187-9
16109747
Biotechniques. 2000 Nov;29(5):948-50, 952
11084852
J Bacteriol. 2003 Mar;185(5):1543-54
12591871
J Bacteriol. 2005 Sep;187(18):6265-72
16159758
FEMS Microbiol Rev. 2008 Jan;32(1):38-55
18070067
Mol Microbiol. 2000 Jan;35(2):435-43
10652104
Mol Microbiol. 1991 Jun;5(6):1469-81
1787797
J Bacteriol. 2002 May;184(9):2500-20
11948165
J Biol Chem. 2007 Jun 22;282(25):18046-18056
17456470
FEMS Microbiol Rev. 1990 Aug;6(4):399-428
2248796
Eur J Biochem. 2002 Jun;269(12):3032-40
12071968
J Bacteriol. 2008 Feb;190(3):1084-96
18039766
Microbiology (Reading). 2003 May;149(Pt 5):1275-1284
12724389
J Bacteriol. 2006 Jan;188(2):659-68
16385055
Mol Microbiol. 1994 Feb;11(3):537-44
8152377
J Bacteriol. 2004 Jul;186(14):4596-604
15231792
J Bacteriol. 2006 Aug;188(15):5494-500
16855239
Mol Cell. 2002 Oct;10(4):779-88
12419222
J Bacteriol. 2007 Jun;189(12):4449-55
17400735
Infect Immun. 2004 Oct;72(10):5638-45
15385461
J Bacteriol. 2007 Mar;189(6):2411-6
17220232
Proc Natl Acad Sci U S A. 2005 May 31;102(22):7823-8
15899978
J Mol Biol. 1996 May 31;259(1):41-57
8648647
Genet Res. 1971 Oct;18(2):185-97
4258431
J Bacteriol. 2001 Sep;183(18):5376-84
11514523
J Bacteriol. 2004 Aug;186(16):5249-57
15292126
Mol Microbiol. 1996 Mar;19(6):1373-84
8730877
J Biol Chem. 1991 Mar 25;266(9):5980-90
2005134
BMC Genomics. 2006 Jun 26;7:162
16800888
J Clin Invest. 2002 Feb;109(3):317-25
11827991
Mol Microbiol. 2008 Jun;68(5):1128-48
18430135
Mol Microbiol. 1998 Aug;29(4):985-97
9767567
Curr Opin Microbiol. 2003 Apr;6(2):140-5
12732303
J Bacteriol. 1984 Dec;160(3):928-34
6438064
J Mol Microbiol Biotechnol. 2001 Oct;3(4):585-600
11545276
Mol Microbiol. 2002 Jan;43(1):107-17
11849540
J Bacteriol. 1995 Jun;177(12):3606-9
7768875
J Bacteriol. 2005 Jul;187(14):4875-83
15995202
Eur J Biochem. 1980 Jan;103(1):117-24
6987054
J Bacteriol. 1991 Mar;173(5):1598-606
1900277
J Bacteriol. 2006 May;188(10):3726-9
16672627
Methods Enzymol. 1994;235:386-405
8057911
Proc Natl Acad Sci U S A. 1993 Dec 1;90(23):11004-8
7504290
Gene. 1998 May 28;212(1):77-86
9661666
Infect Immun. 2007 Nov;75(11):5313-24
17724070
J Bacteriol. 2006 Sep;188(18):6529-38
16952944
Mol Microbiol. 2006 Nov;62(4):1048-63
17078815
J Mol Biol. 2001 Jan 26;305(4):689-702
11162085
J Biol Chem. 2000 May 19;275(20):14795-8
10747855
J Gen Microbiol. 1989 Sep;135(9):2445-51
2516869