Dynamic localization of MreB in Vibrio parahaemolyticus and in the ectopic host bacterium Escherichia coli.
MreB, a homolog of eukaryotic actin, participates in morphogenesis, cell division, cell polarity, and chromosome segregation in prokaryotes. In this study, a yellow fluorescent protein conjugate (YFP-MreB(Vp)) was generated to investigate the behavior of MreB in merodiploid strain SC9 of the enteropathogen Vibrio parahaemolyticus. Under normal growth conditions, YFP-MreB(Vp) formed helical filaments with a pitch of 0.64 +/- 0.09 microm in about 85% of exponential-phase cells, and different clusters, relaxed coils, and ring configurations were observed in a small proportion of the cells. Overexpression of YFP-MreB(Vp) substantially altered the structure of the MreB cytoskeleton and resulted in swollen and pleomorphic cells. Disturbing the activities of penicillin-binding proteins or adding magnesium suppressed the morphological distortions. These results indicate that mislocalization of cell wall-synthesizing machinery was responsible for morphological abnormality. By expressing YFP-MreB(Vp) in the ectopic host bacterium Escherichia coli, shrinkage, fragmentation, and annealing of MreB(Vp) filaments were directly observed. This work revealed the dynamic pattern of the localization of YFP-MreB(Vp) in V. parahaemolyticus and its relationship to cell morphogenesis, and the YFP-MreB(Vp)-E. coli system may be used to investigate the dynamic spatial structures of the MreB cytoskeleton in vivo.
Chiu
Shen-Wen
SW
Department of Microbiology, Soochow University, Taipei, Taiwan, Republic of China.
Chen
Shau-Yan
SY
Wong
Hin-chung
HC
D000199:Actins / Q000378:metabolism*; D019677:Artificial Gene Fusion; D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism*; D004926:Escherichia coli / Q000737:chemistry*; D029968:Escherichia coli Proteins / Q000378:metabolism*; D023061:Gene Order; D017930:Genes, Reporter; D008164:Luminescent Proteins / Q000235:genetics / Q000378:metabolism; D008856:Microscopy, Fluorescence; D005810:Multigene Family; D011993:Recombinant Fusion Proteins / Q000378:metabolism; D026801:Synteny; D014736:Vibrio parahaemolyticus / Q000737:chemistry*
D000199:Actins; D001426:Bacterial Proteins; D029968:Escherichia coli Proteins; D008164:Luminescent Proteins; D011993:Recombinant Fusion Proteins; C054562:yellow fluorescent protein, Bacteria; C078473:MreB protein, E coli
J Bacteriol. 1994 Feb;176(3):917-22
8300545
Nucleic Acids Res. 2001 Jan 1;29(1):123-5
11125067
J Bacteriol. 2006 Feb;188(3):968-76
16428401
J Biol Chem. 2005 Jan 28;280(4):2628-35
15548516
Biosci Biotechnol Biochem. 2006 Nov;70(11):2712-9
17090951
J Bacteriol. 1999 Feb;181(4):1110-7
9973335
Curr Biol. 2007 Feb 6;17(3):266-72
17276920
Cell. 2001 Mar 23;104(6):913-22
11290328
Lancet. 2003 Mar 1;361(9359):743-9
12620739
Proc Natl Acad Sci U S A. 2003 Jun 24;100(13):7865-70
12766229
Mol Microbiol. 2004 Mar;51(5):1321-32
14982627
Mol Microbiol. 2007 Jul;65(1):51-63
17581120
EMBO J. 2002 Jul 1;21(13):3347-57
12093736
Mol Microbiol. 2005 Mar;55(6):1646-57
15752190
J Bacteriol. 1989 Jun;171(6):3123-7
2656641
Appl Environ Microbiol. 2000 Sep;66(9):3981-6
10966418
Proc Natl Acad Sci U S A. 2006 Jul 18;103(29):10929-34
16829583
J Bacteriol. 1998 Apr;180(8):2050-6
9555885
Microbiol Mol Biol Rev. 2006 Sep;70(3):729-54
16959967
Mol Microbiol. 2004 Jul;53(2):469-83
15228528
J Bacteriol. 2005 Jan;187(1):54-64
15601688
Cell. 2003 Jun 13;113(6):767-76
12809607
Proc Natl Acad Sci U S A. 2007 Nov 6;104(45):17795-800
17978175
Microbiol Mol Biol Rev. 2006 Jun;70(2):317-43
16760306
EMBO J. 2002 Jun 17;21(12):3119-27
12065424
EMBO Rep. 2004 Aug;5(8):789-94
15272301
Plasmid. 2004 May;51(3):246-55
15109831
J Bacteriol. 2007 Oct;189(20):7450-63
17704222
J Bacteriol. 1998 Sep;180(17):4555-63
9721295
Proc Natl Acad Sci U S A. 2004 Jun 8;101(23):8643-8
15159537
Science. 2004 Nov 5;306(5698):1021-5
15528442
Mol Microbiol. 2005 Sep;57(5):1196-209
16101995
Mol Microbiol. 2006 May;60(4):838-52
16677297
EMBO J. 2003 Oct 1;22(19):5283-92
14517265
Dev Cell. 2006 Sep;11(3):399-409
16950129
Proc Natl Acad Sci U S A. 2005 Dec 6;102(49):17658-63
16306264
Dev Cell. 2003 Jan;4(1):19-28
12530960
Mol Microbiol. 1999 Feb;31(4):1149-59
10096082
J Bacteriol. 1993 May;175(10):3139-45
8387996
Antimicrob Agents Chemother. 1999 Nov;43(11):2702-9
10543750