title: Polyamines are required for the expression of key Hms proteins important for Yersinia pestis biofilm formation.
abstract: We previously showed that mutations in the genes encoding the two main biosynthetic enzymes responsible for polyamine production, arginine decarboxylase (SpeA) and ornithine decarboxylase (SpeC) cause a loss of biofilm formation in Yersinia pestis. In Y. pestis the development of a biofilm is dependent on 6 Hms (hemin storage) proteins (HmsH, F, R, S, T and P) grouped into 3 operons; hmsHFRS, hmsT and hmsP. In this article we show that polyamines are necessary to maintain the levels of key Hms proteins. In the absence of polyamines there is an approximately 93%, approximately 43% and approximately 90% reduction in protein levels of HmsR, HmsS and HmsT respectively. Overexpression of hmsR and hmsT from plasmids alone can restore biofilm formation to a SpeA(-)SpeC(-) mutant. Addition of exogenous putrescine also restores normal levels of HmsR, HmsS, HmsT and biofilm production. Analyses using transcriptional reporters and quantitative RT-PCR indicate that the initiation of transcription and mRNA stability are not reduced by polyamine deficiency. Instead, translational reporters indicate that polyamines function at least in part by modulating the translation of HmsR and HmsT. Although construction of a consensus Shine-Dalgarno sequence upstream of hmsT modestly reduced the stimulation of translation by putrescine, additional mechanisms likely contribute to the polyamine-dependent expression of HmsT. Finally, we have shown that polyamines play a role in bubonic plague.
authors.affiliation: Department of Pharmaceutical Sciences, College of Pharmacy, University of Kentucky, Lexington, KY 40536, USA.
authors.forename: Brian W
authors.initials: BW
authors.lastname: Wortham
authors.forename: Marcos A
authors.initials: MA
authors.lastname: Oliveira
authors.forename: Jacqueline D
authors.initials: JD
authors.lastname: Fetherston
authors.forename: Robert D
authors.initials: RD
authors.lastname: Perry
mesh_terms: D000818:Animals; D001426:Bacterial Proteins / Q000096:biosynthesis*; D018441:Biofilms / Q000254:growth & development*; D004195:Disease Models, Animal; D020869:Gene Expression Profiling; D015964:Gene Expression Regulation, Bacterial / Q000187:drug effects*; D017930:Genes, Reporter; D051379:Mice; D010930:Plague / Q000382:microbiology; D011073:Polyamines / Q000378:metabolism*; D020871:RNA Stability; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014774:Virulence; D015010:Yersinia pestis / Q000254:growth & development / Q000378:metabolism / Q000472:pathogenicity / Q000502:physiology*; D001616:beta-Galactosidase / Q000235:genetics / Q000378:metabolism
chemical_list: D001426:Bacterial Proteins; C122178:HMST protein, Yersinia pestis; D011073:Polyamines; D001616:beta-Galactosidase
references.citation: Mol Cell Biochem. 1991 Feb 2;100(2):129-40
references.pmid: 2008175
references.citation: Microbiology (Reading). 2006 Nov;152(Pt 11):3399-3410
references.pmid: 17074909
references.citation: J Bacteriol. 1961 Apr;81:605-8
references.pmid: 13714203
references.citation: J Chromatogr B Analyt Technol Biomed Life Sci. 2002 Dec 5;781(1-2):107-49
references.pmid: 12450656
references.citation: Mol Microbiol. 2004 Oct;54(1):75-88
references.pmid: 15458406
references.citation: Mol Microbiol. 1992 Sep;6(18):2693-704
references.pmid: 1447977
references.citation: J Bacteriol. 2005 Jan;187(1):382-7
references.pmid: 15601723
references.citation: J Biochem. 2006 Jan;139(1):11-6
references.pmid: 16428314
references.citation: Trends Microbiol. 2001 Jan;9(1):34-9
references.pmid: 11166241
references.citation: Adv Exp Med Biol. 2007;603:192-200
references.pmid: 17966415
references.citation: J Biol Chem. 2009 Apr 10;284(15):9899-907
references.pmid: 19196710
references.citation: Nature. 1975 Mar 6;254(5495):34-8
references.pmid: 803646
references.citation: Curr Opin Microbiol. 2007 Feb;10(1):17-23
references.pmid: 17208514
references.citation: Methods Cell Biol. 1991;34:61-75
references.pmid: 1943817
references.citation: Clin Chem. 1979 Sep;25(9):1564-71
references.pmid: 466783
references.citation: Infect Immun. 2001 May;69(5):2829-37
references.pmid: 11292695
references.citation: Methods Mol Biol. 2000;132:365-86
references.pmid: 10547847
references.citation: J Bacteriol. 1978 Jun;134(3):1141-56
references.pmid: 149110
references.citation: Infect Immun. 1999 Sep;67(9):4801-13
references.pmid: 10456934
references.citation: Mol Microbiol. 1999 Apr;32(2):403-14
references.pmid: 10231495
references.citation: Microbiology (Reading). 1999 Jan;145 ( Pt 1):197-209
references.pmid: 10206699
references.citation: J Biol Chem. 1999 Aug 6;274(32):22723-8
references.pmid: 10428855
references.citation: J Biol Chem. 2004 Oct 29;279(44):46008-13
references.pmid: 15326188
references.citation: Nat Rev Microbiol. 2004 Feb;2(2):95-108
references.pmid: 15040259
references.citation: Microbiology (Reading). 2007 Jul;153(Pt 7):2332-2341
references.pmid: 17600077
references.citation: Biochem J. 1999 Dec 15;344 Pt 3:633-42
references.pmid: 10585849
references.citation: FEMS Microbiol Lett. 2005 Jun 15;247(2):123-30
references.pmid: 15935569
references.citation: Eur Biophys J. 2006 Feb;35(3):214-23
references.pmid: 16247626
references.citation: J Biol Chem. 2006 Apr 7;281(14):9527-37
references.pmid: 16476727
references.citation: Appl Microbiol. 1969 Nov;18(5):834-7
references.pmid: 5370459
references.citation: J Biol Chem. 1997 Feb 14;272(7):4058-64
references.pmid: 9020114
references.citation: BMC Biochem. 2007 Jan 15;8:1
references.pmid: 17224065
references.citation: J Bacteriol. 2004 May;186(9):2724-34
references.pmid: 15090514
references.citation: Science. 2006 Feb 24;311(5764):1113-6
references.pmid: 16497924
references.citation: Methods Enzymol. 1999;310:91-109
references.pmid: 10547784
references.citation: FEMS Microbiol Lett. 2004 Jul 15;236(2):163-73
references.pmid: 15251193
references.citation: J Infect Dis. 2004 Aug 15;190(4):783-92
references.pmid: 15272407
references.citation: Arch Biochem Biophys. 1993 Jan;300(1):63-8
references.pmid: 7678729
references.citation: J Bacteriol. 2005 Nov;187(21):7434-43
references.pmid: 16237027
references.citation: J Bacteriol. 1991 Jan;173(2):417-25
references.pmid: 1898928
references.citation: Nucleic Acids Res. 2001 May 1;29(9):e45
references.pmid: 11328886
references.citation: Environ Microbiol. 2008 Jun;10(6):1419-32
references.pmid: 18279344
references.citation: J Bacteriol. 2005 Oct;187(19):6816-23
references.pmid: 16166544
references.citation: Methods. 2001 Dec;25(4):402-8
references.pmid: 11846609
references.citation: Gene. 1997 Jul 1;193(1):13-21
references.pmid: 9249062
references.citation: Philos Trans R Soc Lond B Biol Sci. 2007 Jul 29;362(1483):1119-34
references.pmid: 17360280
references.citation: Microbiol Mol Biol Rev. 2009 Jun;73(2):310-47
references.pmid: 19487730
references.citation: Science. 1996 Jul 19;273(5273):367-70
references.pmid: 8662526
references.citation: Microbiology (Reading). 2010 May;156(Pt 5):1424-1438
references.pmid: 20093287
references.citation: Microbiology (Reading). 1999 Aug;145 ( Pt 8):2117-2128
references.pmid: 10463178
references.citation: J Bacteriol. 2004 Mar;186(6):1638-47
references.pmid: 14996794
references.citation: Proc Natl Acad Sci U S A. 1979 Sep;76(9):4350-4
references.pmid: 388439
references.citation: EMBO Rep. 2005 Oct;6(10):992-7
references.pmid: 16170309
references.citation: J Bacteriol. 2006 Apr;188(7):2355-63
references.pmid: 16547021
references.citation: Nucleic Acids Res. 1979 Nov 24;7(6):1513-23
references.pmid: 388356
references.citation: J Bacteriol. 2009 Sep;191(17):5348-57
references.pmid: 19542278
references.citation: J Bacteriol. 2002 Jul;184(14):3765-73
references.pmid: 12081945
references.citation: Biometals. 2007 Jun;20(3-4):699-703
references.pmid: 17206386
references.citation: Gene. 1991 Dec 20;109(1):137-41
references.pmid: 1756974
references.citation: J Bacteriol. 1994 Sep;176(17):5385-92
references.pmid: 7915269
references.citation: FEMS Microbiol Lett. 2009 Oct;299(2):166-74
references.pmid: 19694812
references.citation: Br J Exp Pathol. 1956 Dec;37(6):570-6
references.pmid: 13396141
references.citation: Annu Rev Genet. 2006;40:385-407
references.pmid: 16895465
references.citation: J Bacteriol. 2007 Mar;189(6):2359-68
references.pmid: 17220219
references.citation: J Biol Chem. 1998 Jul 10;273(28):17604-9
references.pmid: 9651355
references.citation: Nucleic Acids Res. 2005 Mar 23;33(6):1790-803
references.pmid: 15788751
references.citation: J Bacteriol. 1995 Apr;177(7):1824-33
references.pmid: 7896707
references.citation: J Bacteriol. 1995 Jul;177(14):4121-30
references.pmid: 7608087