Interdomain communication in the Mycobacterium tuberculosis environmental phosphatase Rv1364c.
An "environmental phosphatase" controls bacterial transcriptional responses through alternative sigma factor subunits of RNA polymerase and a partner switching mechanism has been proposed to mediate phosphatase regulation. In many bacteria, the environmental phosphatase and multiple regulators are encoded in separate genes whose products form transient complexes. In contrast, in the Mycobacterium tuberculosis homolog, Rv1364c, the phosphatase is fused to two characteristic regulatory modules with sequence similarities to anti-sigma factor kinases and anti-anti-sigma factor proteins. Here we exploit this fusion to explore interactions between the phosphatase and the regulatory domains. We show quantitatively that the anti-sigma factor kinase domain activates the phosphatase domain, the kinase-phosphatase fusion protein autophosphorylates in Escherichia coli, and phosphorylation is antagonized by the phosphatase activity. Small angle x-ray scattering defines solution structures consistent with the interdomain communication observed biochemically. Taken together, these data indicate that Rv1364c provides a single chain framework to understand the structure, function, and regulation of environmental phosphatases throughout the bacterial kingdom.
Greenstein
Andrew E
AE
Department of Molecular and Cell Biology, University of California, Berkeley, California 94720-3220, USA.
Hammel
Michal
M
Cavazos
Alexandra
A
Alber
Tom
T
D009169:Mycobacterium tuberculosis / Q000201:enzymology*; D010744:Phosphoric Monoester Hydrolases / Q000737:chemistry* / Q000378:metabolism; D010766:Phosphorylation / Q000502:physiology; D017434:Protein Structure, Tertiary / Q000502:physiology; D012542:Scattering, Radiation; D013329:Structure-Activity Relationship; D014965:X-Rays
D010744:Phosphoric Monoester Hydrolases
Tuberculosis (Edinb). 2004;84(3-4):228-38
15207492
FEBS J. 2008 Dec;275(24):6295-308
19016841
J Mol Biol. 2004 Sep 24;342(4):1187-95
15351644
Mol Microbiol. 2003 Nov;50(3):949-59
14617153
FEMS Microbiol Rev. 2006 Nov;30(6):926-41
17064287
Biophys J. 2001 Jun;80(6):2946-53
11371467
J Bacteriol. 2002 Sep;184(17):4881-90
12169614
Proc Natl Acad Sci U S A. 2002 Feb 19;99(4):1831-5
11830637
Infect Immun. 2004 Mar;72(3):1733-45
14977982
Front Biosci. 2004 May 01;9:1136-56
14977534
Mol Microbiol. 2006 Dec;62(5):1251-63
17064366
J Bacteriol. 2004 Oct;186(20):6830-6
15466036
Protein Sci. 1995 Aug;4(8):1596-607
8520486
Annu Rev Microbiol. 1998;52:231-86
9891799
J Struct Biol. 1999 Apr-May;125(2-3):185-95
10222274
Mol Microbiol. 2002 Sep;45(6):1527-40
12354223
Tuberculosis (Edinb). 2005 Sep-Nov;85(5-6):347-55
16263329
Q Rev Biophys. 2007 Aug;40(3):191-285
18078545
J Mol Microbiol Biotechnol. 2005;9(2):65-76
16319496
J Mol Biol. 2004 Jul 30;341(1):135-50
15312768
Science. 1999 Jul 30;285(5428):751-3
10427000
Proteins. 2008 Feb 15;70(3):611-25
17876813
J Biol Chem. 1999 Jul 16;274(29):20336-43
10400656
J Bacteriol. 2002 Oct;184(20):5583-9
12270815
Infect Genet Evol. 2004 Dec;4(4):301-8
15374527
J Bacteriol. 2004 Sep;186(18):6124-32
15342582
Anal Biochem. 2005 Aug 15;343(2):313-21
15993367
Mol Microbiol. 2003 Sep;49(6):1657-69
12950928
Nat Methods. 2009 Aug;6(8):606-12
19620974
Mol Microbiol. 2000 Jan;35(1):180-8
10632888
Genes Dev. 1996 Sep 15;10(18):2265-75
8824586
PLoS Pathog. 2007 Apr;3(4):e49
17411339
Curr Opin Microbiol. 2008 Apr;11(2):121-7
18375176