Identification and function of the RNA chaperone Hfq in the Lyme disease spirochete Borrelia burgdorferi.
Hfq is a global regulatory RNA-binding protein. We have identified and characterized an atypical Hfq required for gene regulation and infectivity in the Lyme disease spirochete Borrelia burgdorferi. Sequence analyses of the putative B. burgdorferi Hfq protein revealed only a modest level of similarity with the Hfq from Escherichia coli, although a few key residues are retained and the predicted tertiary structure is similar. Several lines of evidence suggest that the B. burgdorferi bb0268 gene encodes a functional Hfq homologue. First, the hfq(Bb) gene (bb0268) restores the efficient translation of an rpoS::lacZ fusion in an E. coli hfq null mutant. Second, the Hfq from B. burgdorferi binds to the small RNA DsrA(Bb) and the rpoS mRNA. Third, a B. burgdorferi hfq null mutant was generated and has a pleiotropic phenotype that includes increased cell length and decreased growth rate, as found in hfq mutants in other bacteria. The hfq(Bb) mutant phenotype is complemented in trans with the hfq gene from either B. burgdorferi or, surprisingly, E. coli. This is the first example of a heterologous bacterial gene complementing a B. burgdorferi mutant. The alternative sigma factor RpoS and the outer membrane lipoprotein OspC, which are induced by increased temperature and required for mammalian infection, are not upregulated in the hfq mutant. Consequently, the hfq mutant is not infectious by needle inoculation in the murine model. These data suggest that Hfq plays a key role in the regulation of pathogenicity factors in B. burgdorferi and we hypothesize that the spirochete has a complex Hfq-dependent sRNA network.
Lybecker
Meghan C
MC
Division of Biological Sciences, The University of Montana, Missoula, MT 59812-4824, USA.
Abel
Cassandra A
CA
Feig
Andrew L
AL
Samuels
D Scott
DS
D000595:Amino Acid Sequence; D000818:Animals; D001426:Bacterial Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D025065:Borrelia burgdorferi / Q000737:chemistry / Q000235:genetics / Q000378:metabolism* / Q000472:pathogenicity; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D029968:Escherichia coli Proteins / Q000235:genetics / Q000378:metabolism; D005260:Female; D015964:Gene Expression Regulation, Bacterial; D005816:Genetic Complementation Test; D035001:Host Factor 1 Protein / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D006801:Humans; D008193:Lyme Disease / Q000382:microbiology*; D051379:Mice; D008809:Mice, Inbred C3H; D018832:Molecular Chaperones / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D008969:Molecular Sequence Data; D016601:RNA-Binding Proteins / Q000235:genetics / Q000378:metabolism; D016415:Sequence Alignment; D014774:Virulence
D001426:Bacterial Proteins; D029968:Escherichia coli Proteins; C087373:Hfq protein, E coli; D035001:Host Factor 1 Protein; D018832:Molecular Chaperones; D016601:RNA-Binding Proteins
Mol Microbiol. 2007 Jan;63(1):193-217
17163975
Curr Opin Microbiol. 2006 Apr;9(2):229-36
16529986
Med Microbiol Immunol. 2000 Nov;189(2):59-66
11138638
FEMS Microbiol Rev. 2006 Jan;30(1):3-16
16438677
J Bacteriol. 2003 Nov;185(22):6723-7
14594849
Nucleic Acids Res. 2003 Jul 15;31(14):4091-8
12853626
J Bacteriol. 1996 Mar;178(6):1607-13
8626288
J Bacteriol. 1997 Dec;179(23):7476-87
9393714
Nat Protoc. 2009;4(3):363-71
19247286
Annu Rev Entomol. 1991;36:587-609
2006870
Ann N Y Acad Sci. 1994 Dec 15;740:146-56
7840446
Nature. 1997 Dec 11;390(6660):580-6
9403685
Nat Rev Genet. 2007 Oct;8(10):776-90
17846637
EMBO J. 1999 Oct 15;18(20):5789-802
10523320
Mol Microbiol. 2007 Feb;63(4):1259-73
17257307
Nucleic Acids Res. 2003 Dec 15;31(24):7302-10
14654705
J Comput Chem. 2004 Oct;25(13):1605-12
15264254
Cell. 2009 Feb 20;136(4):615-28
19239884
Nature. 1968 Aug 10;219(5154):588-90
4874917
Proc Natl Acad Sci U S A. 1995 Mar 28;92(7):2909-13
7708747
Methods Mol Biol. 1995;47:253-9
7550741
J Biol Chem. 1995 May 12;270(19):11181-9
7538113
Genes Dev. 2000 May 1;14(9):1109-18
10809669
J Bacteriol. 2009 Apr;191(8):2902-5
19201806
Curr Opin Microbiol. 2007 Apr;10(2):125-33
17395525
J Biol Chem. 2005 Mar 4;280(9):7901-8
15618228
RNA. 2005 Jun;11(6):976-84
15872186
Infect Immun. 2003 Apr;71(4):1689-705
12654782
RNA. 2008 Mar;14(3):514-23
18230766
Cell. 1999 Feb 5;96(3):375-87
10025403
Annu Rev Microbiol. 2004;58:303-28
15487940
Curr Opin Microbiol. 2010 Feb;13(1):24-33
20080057
EMBO J. 2002 Jul 1;21(13):3546-56
12093755
J Mol Biol. 2002 Jun 28;320(1):129-42
12079339
Mol Microbiol. 2004 Mar;51(6):1525-33
15009882
Mol Microbiol. 2003 Jun;48(6):1665-77
12791146
N Engl J Med. 1983 Mar 31;308(13):733-40
6828118
Proc Natl Acad Sci U S A. 2000 Dec 5;97(25):13865-70
11106398
J Mol Biol. 2002 Jul 19;320(4):705-12
12095248
EMBO J. 2004 Jan 28;23(2):396-405
14739933
Mol Microbiol. 2007 May;64(4):1075-89
17501929
RNA. 2003 Jan;9(1):33-43
12554874
Trends Biochem Sci. 2000 Mar;25(3):147-50
10694887
Genes Dev. 2003 Oct 1;17(19):2374-83
12975324
Nucleic Acids Res. 2002 Sep 1;30(17):3662-71
12202750
Infect Immun. 2001 Jan;69(1):446-55
11119536
N Engl J Med. 1983 Mar 31;308(13):740-2
6828119
Nucleic Acids Res. 2003 Jul 1;31(13):3406-15
12824337
Proc Natl Acad Sci U S A. 2003 Sep 16;100(19):11001-6
12949258
EMBO Rep. 2003 Mar;4(3):284-9
12634847
J Bacteriol. 1997 Feb;179(3):656-62
9006017
Mol Microbiol. 2009 Dec;74(6):1295-9
19943896
J Bacteriol. 2005 Jul;187(14):4822-9
15995197
Mol Microbiol. 2000 Nov;38(4):667-72
11115103
Proc Natl Acad Sci U S A. 2002 Apr 2;99(7):4620-5
11917098
EMBO J. 2001 May 1;20(9):2293-303
11331594
Mol Microbiol. 1994 Jul;13(1):35-49
7984093
Mol Microbiol. 2003 May;48(4):855-61
12753181
Mol Microbiol. 2006 Aug;61(4):1013-22
16859494
Acta Crystallogr Sect F Struct Biol Cryst Commun. 2010 Jul 1;66(Pt 7):760-4
20606268
Proc Natl Acad Sci U S A. 2004 May 11;101(19):7293-8
15123801
Crit Rev Biochem Mol Biol. 2005 Mar-Apr;40(2):93-113
15814430
Proc Natl Acad Sci U S A. 2002 Feb 5;99(3):1562-7
11830671
Mol Microbiol. 2007 Sep;65(5):1193-217
17645733
J Bacteriol. 1997 Jan;179(1):297-300
8982015
Infect Immun. 1995 Nov;63(11):4535-9
7591099
Proc Natl Acad Sci U S A. 2001 Mar 27;98(7):3685-9
11259661
Curr Opin Microbiol. 2009 Dec;12(6):674-82
19880344
J Bacteriol. 2001 Mar;183(6):1997-2005
11222598
J Mol Biol. 2004 Dec 10;344(5):1211-23
15561140
Proc Natl Acad Sci U S A. 2001 Oct 23;98(22):12724-9
11675503
Mol Microbiol. 2007 Jul;65(2):277-93
17590233
Yale J Biol Med. 1984 Jul-Aug;57(4):521-5
6393604
Curr Opin Microbiol. 2003 Apr;6(2):120-4
12732300
Infect Immun. 2004 Nov;72(11):6433-45
15501774
Mol Microbiol. 2000 Sep;37(6):1470-9
10998177
Lancet Infect Dis. 2004 Sep;4(9):575-83
15336225
J Exp Med. 2004 Mar 1;199(5):641-8
14981112
Nat Struct Mol Biol. 2004 Dec;11(12):1206-14
15531892
Microbiol Mol Biol Rev. 2002 Sep;66(3):373-95, table of contents
12208995
Mol Microbiol. 2004 Jul;53(1):345-54
15225327
Proc Natl Acad Sci U S A. 2009 Nov 17;106(46):19292-7
19889981
Mol Cell. 2002 Jan;9(1):23-30
11804583
Cell Mol Life Sci. 2009 Aug;66(16):2661-76
19554260
J Bacteriol. 2000 May;182(9):2445-52
10762244
RNA. 2003 Nov;9(11):1308-14
14561880
Science. 1982 Jun 18;216(4552):1317-9
7043737
Mol Microbiol. 1999 Nov;34(4):690-700
10564509
Curr Opin Microbiol. 2004 Apr;7(2):140-4
15063850
Mol Cell. 2002 Jan;9(1):11-22
11804582
RNA. 2007 Dec;13(12):2213-23
17959927
EMBO J. 2007 Feb 21;26(4):965-75
17268550