title: Phosphorylation of PhoP protein plays direct regulatory role in lipid biosynthesis of Mycobacterium tuberculosis.
abstract: Mycobacterium tuberculosis PhoP is essential for virulence and intracellular growth of the tubercle bacilli. Genetic evidence suggests that PhoP regulates complex lipid biosynthesis, and absence of some of these lipid molecules in a phoP mutant partly accounts for its attenuated growth in macrophages and/or mice. To investigate the mechanism of regulation, here we demonstrate the essentiality of phosphorylation of PhoP in the regulation of complex lipid biosynthesis. We show that phosphorylated PhoP activates transcription of pks2 and msl3, gene(s) encoding polyketide β-ketoacyl synthases through direct DNA binding at the upstream regulatory region(s) of the target genes. Our results identify the genetic determinants recognized by PhoP and show that activation of target genes requires interaction(s) of the phosphorylated regulator at the cognate binding sites. The fact that these sites within the regulatory region of respective genes do not bind in vitro with either unphosphorylated or phosphorylation-deficient PhoP protein is consistent with phosphorylation-dependent assembly of the transcription initiation complex leading to in vivo transcriptional activation. Together, these results reveal so far unknown molecular mechanisms of how PhoP contributes to M. tuberculosis cell wall composition by regulating complex lipid biosynthesis.
authors.affiliation: Institute of Microbial Technology, Council of Scientific and Industrial Research, Sector 39 A, Chandigarh 160036, India.
authors.forename: Rajni
authors.initials: R
authors.lastname: Goyal
authors.forename: Arijit Kumar
authors.initials: AK
authors.lastname: Das
authors.forename: Ranjeet
authors.initials: R
authors.lastname: Singh
authors.forename: Pradip K
authors.initials: PK
authors.lastname: Singh
authors.forename: Suresh
authors.initials: S
authors.lastname: Korpole
authors.forename: Dibyendu
authors.initials: D
authors.lastname: Sarkar
mesh_terms: D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism*; D002473:Cell Wall / Q000235:genetics / Q000378:metabolism*; D064429:Fatty Acid Synthases / Q000235:genetics / Q000378:metabolism; D008055:Lipids / Q000096:biosynthesis* / Q000235:genetics; D009169:Mycobacterium tuberculosis / Q000235:genetics / Q000378:metabolism*; D010766:Phosphorylation / Q000502:physiology; D020218:Response Elements / Q000502:physiology*; D014158:Transcription, Genetic / Q000502:physiology*
chemical_list: D001426:Bacterial Proteins; D008055:Lipids; C061064:PhoP protein, Bacteria; D064429:Fatty Acid Synthases; C429525:Pks2 protein, Mycobacterium tuberculosis
references.citation: J Bacteriol. 1999 Jan;181(2):469-76
references.pmid: 9882660
references.citation: Microbiology (Reading). 2002 Oct;148(Pt 10):3029-3037
references.pmid: 12368436
references.citation: Mol Microbiol. 2001 Jul;41(1):179-87
references.pmid: 11454210
references.citation: FEBS Lett. 2006 Oct 2;580(22):5328-38
references.pmid: 16979633
references.citation: Biochemistry. 2007 Dec 25;46(51):14751-61
references.pmid: 18052041
references.citation: PLoS One. 2010 Jun 10;5(6):e11066
references.pmid: 20548782
references.citation: Proc Natl Acad Sci U S A. 2001 Jun 19;98(13):7534-9
references.pmid: 11416222
references.citation: Curr Opin Microbiol. 2010 Apr;13(2):160-7
references.pmid: 20080056
references.citation: J Bacteriol. 1994 Nov;176(21):6749-53
references.pmid: 7961429
references.citation: J Mol Biol. 1986 May 5;189(1):113-30
references.pmid: 3537305
references.citation: J Bacteriol. 2009 Dec;191(24):7466-76
references.pmid: 19820095
references.citation: Structure. 2002 May;10(5):701-13
references.pmid: 12015152
references.citation: Proc Natl Acad Sci U S A. 2003 May 13;100(10):6133-8
references.pmid: 12730372
references.citation: Mol Microbiol. 2002 Sep;45(5):1451-9
references.pmid: 12207710
references.citation: Cell Host Microbe. 2008 Feb 14;3(2):97-103
references.pmid: 18312844
references.citation: J Bacteriol. 2008 Feb;190(4):1317-28
references.pmid: 18065544
references.citation: Vaccine. 2006 Apr 24;24(17):3408-19
references.pmid: 16564606
references.citation: PLoS Pathog. 2008 Feb 8;4(2):e33
references.pmid: 18282096
references.citation: Methods Mol Biol. 1998;101:91-107
references.pmid: 9921472
references.citation: PLoS One. 2008;3(10):e3496
references.pmid: 18946503
references.citation: Mol Microbiol. 2002 Feb;43(3):717-31
references.pmid: 11929527
references.citation: J Biol Chem. 2006 Apr 7;281(14):9659-66
references.pmid: 16434396
references.citation: Mol Microbiol. 1999 Jan;31(2):715-24
references.pmid: 10027986
references.citation: Mol Microbiol. 2011 May;80(3):678-94
references.pmid: 21401735
references.citation: J Biol Chem. 2010 Nov 5;285(45):34309-18
references.pmid: 20814030
references.citation: Trends Biochem Sci. 2007 May;32(5):225-34
references.pmid: 17433693
references.citation: Infect Immun. 2003 Aug;71(8):4684-90
references.pmid: 12874349
references.citation: Biochemistry. 2011 Jul 5;50(26):5948-57
references.pmid: 21634789
references.citation: J Bacteriol. 2008 Feb;190(4):1329-34
references.pmid: 18065542
references.citation: Microbiology (Reading). 2004 Jan;150(Pt 1):241-246
references.pmid: 14702417
references.citation: Proc Natl Acad Sci U S A. 2003 May 13;100(10):6121-6
references.pmid: 12724526
references.citation: Trends Microbiol. 2008 Nov;16(11):528-34
references.pmid: 18835713
references.citation: Mol Cell. 2000 Apr;5(4):717-27
references.pmid: 10882107
references.citation: J Bacteriol. 2005 Jan;187(1):202-12
references.pmid: 15601704
references.citation: Anal Biochem. 2009 Jun 1;389(1):77-9
references.pmid: 19302974
references.citation: J Bacteriol. 1999 Jun;181(11):3402-8
references.pmid: 10348851
references.citation: Nucleic Acids Res. 2005 Feb 01;33(2):e21
references.pmid: 15687379
references.citation: J Biol Chem. 2006 Jan 20;281(3):1313-6
references.pmid: 16326699
references.citation: Cell Microbiol. 2003 Jun;5(6):405-15
references.pmid: 12780778
references.citation: J Exp Med. 2003 Sep 1;198(5):705-13
references.pmid: 12953092
references.citation: Mol Microbiol. 1990 Nov;4(11):1911-9
references.pmid: 2082148
references.citation: Structure. 2002 Aug;10(8):1041-50
references.pmid: 12176382
references.citation: PLoS Pathog. 2010 Sep 09;6(9):e1001100
references.pmid: 20844580
references.citation: Proc Natl Acad Sci U S A. 2001 Oct 23;98(22):12706-11
references.pmid: 11675502
references.citation: J Biol Chem. 2004 May 14;279(20):21257-65
references.pmid: 15001577
references.citation: J Bacteriol. 2004 Apr;186(8):2476-80
references.pmid: 15060051
references.citation: PLoS One. 2010 Sep 24;5(9):e12978
references.pmid: 20885976
references.citation: J Bacteriol. 2004 Aug;186(16):5197-201
references.pmid: 15292120
references.citation: Trends Microbiol. 2003 Aug;11(8):359-66
references.pmid: 12915093
references.citation: Proc Natl Acad Sci U S A. 1999 Sep 28;96(20):11554-9
references.pmid: 10500215
references.citation: J Biol Chem. 2001 May 18;276(20):16833-9
references.pmid: 11278910
references.citation: Mol Microbiol. 2006 Apr;60(2):312-30
references.pmid: 16573683