Polyamines are required for the expression of key Hms proteins important for Yersinia pestis biofilm formation.
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.
Wortham
Brian W
BW
Department of Pharmaceutical Sciences, College of Pharmacy, University of Kentucky, Lexington, KY 40536, USA.
Oliveira
Marcos A
MA
Fetherston
Jacqueline D
JD
Perry
Robert D
RD
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
D001426:Bacterial Proteins; C122178:HMST protein, Yersinia pestis; D011073:Polyamines; D001616:beta-Galactosidase
Mol Cell Biochem. 1991 Feb 2;100(2):129-40
2008175
Microbiology (Reading). 2006 Nov;152(Pt 11):3399-3410
17074909
J Bacteriol. 1961 Apr;81:605-8
13714203
J Chromatogr B Analyt Technol Biomed Life Sci. 2002 Dec 5;781(1-2):107-49
12450656
Mol Microbiol. 2004 Oct;54(1):75-88
15458406
Mol Microbiol. 1992 Sep;6(18):2693-704
1447977
J Bacteriol. 2005 Jan;187(1):382-7
15601723
J Biochem. 2006 Jan;139(1):11-6
16428314
Trends Microbiol. 2001 Jan;9(1):34-9
11166241
Adv Exp Med Biol. 2007;603:192-200
17966415
J Biol Chem. 2009 Apr 10;284(15):9899-907
19196710
Nature. 1975 Mar 6;254(5495):34-8
803646
Curr Opin Microbiol. 2007 Feb;10(1):17-23
17208514
Methods Cell Biol. 1991;34:61-75
1943817
Clin Chem. 1979 Sep;25(9):1564-71
466783
Infect Immun. 2001 May;69(5):2829-37
11292695
Methods Mol Biol. 2000;132:365-86
10547847
J Bacteriol. 1978 Jun;134(3):1141-56
149110
Infect Immun. 1999 Sep;67(9):4801-13
10456934
Mol Microbiol. 1999 Apr;32(2):403-14
10231495
Microbiology (Reading). 1999 Jan;145 ( Pt 1):197-209
10206699
J Biol Chem. 1999 Aug 6;274(32):22723-8
10428855
J Biol Chem. 2004 Oct 29;279(44):46008-13
15326188
Nat Rev Microbiol. 2004 Feb;2(2):95-108
15040259
Microbiology (Reading). 2007 Jul;153(Pt 7):2332-2341
17600077
Biochem J. 1999 Dec 15;344 Pt 3:633-42
10585849
FEMS Microbiol Lett. 2005 Jun 15;247(2):123-30
15935569
Eur Biophys J. 2006 Feb;35(3):214-23
16247626
J Biol Chem. 2006 Apr 7;281(14):9527-37
16476727
Appl Microbiol. 1969 Nov;18(5):834-7
5370459
J Biol Chem. 1997 Feb 14;272(7):4058-64
9020114
BMC Biochem. 2007 Jan 15;8:1
17224065
J Bacteriol. 2004 May;186(9):2724-34
15090514
Science. 2006 Feb 24;311(5764):1113-6
16497924
Methods Enzymol. 1999;310:91-109
10547784
FEMS Microbiol Lett. 2004 Jul 15;236(2):163-73
15251193
J Infect Dis. 2004 Aug 15;190(4):783-92
15272407
Arch Biochem Biophys. 1993 Jan;300(1):63-8
7678729
J Bacteriol. 2005 Nov;187(21):7434-43
16237027
J Bacteriol. 1991 Jan;173(2):417-25
1898928
Nucleic Acids Res. 2001 May 1;29(9):e45
11328886
Environ Microbiol. 2008 Jun;10(6):1419-32
18279344
J Bacteriol. 2005 Oct;187(19):6816-23
16166544
Methods. 2001 Dec;25(4):402-8
11846609
Gene. 1997 Jul 1;193(1):13-21
9249062
Philos Trans R Soc Lond B Biol Sci. 2007 Jul 29;362(1483):1119-34
17360280
Microbiol Mol Biol Rev. 2009 Jun;73(2):310-47
19487730
Science. 1996 Jul 19;273(5273):367-70
8662526
Microbiology (Reading). 2010 May;156(Pt 5):1424-1438
20093287
Microbiology (Reading). 1999 Aug;145 ( Pt 8):2117-2128
10463178
J Bacteriol. 2004 Mar;186(6):1638-47
14996794
Proc Natl Acad Sci U S A. 1979 Sep;76(9):4350-4
388439
EMBO Rep. 2005 Oct;6(10):992-7
16170309
J Bacteriol. 2006 Apr;188(7):2355-63
16547021
Nucleic Acids Res. 1979 Nov 24;7(6):1513-23
388356
J Bacteriol. 2009 Sep;191(17):5348-57
19542278
J Bacteriol. 2002 Jul;184(14):3765-73
12081945
Biometals. 2007 Jun;20(3-4):699-703
17206386
Gene. 1991 Dec 20;109(1):137-41
1756974
J Bacteriol. 1994 Sep;176(17):5385-92
7915269
FEMS Microbiol Lett. 2009 Oct;299(2):166-74
19694812
Br J Exp Pathol. 1956 Dec;37(6):570-6
13396141
Annu Rev Genet. 2006;40:385-407
16895465
J Bacteriol. 2007 Mar;189(6):2359-68
17220219
J Biol Chem. 1998 Jul 10;273(28):17604-9
9651355
Nucleic Acids Res. 2005 Mar 23;33(6):1790-803
15788751
J Bacteriol. 1995 Apr;177(7):1824-33
7896707
J Bacteriol. 1995 Jul;177(14):4121-30
7608087