title: A role for FtsA in SPOR-independent localization of the essential Escherichia coli cell division protein FtsN.
abstract: FtsN is a bitopic membrane protein and the last essential component to localize to the Escherichia coli cell division machinery, or divisome. The periplasmic SPOR domain of FtsN was previously shown to localize to the divisome in a self-enhancing manner, relying on the essential activity of FtsN and the peptidoglycan synthesis and degradation activities of FtsI and amidases respectively. Because FtsN has a known role in recruiting amidases and is predicted to stimulate the activity of FtsI, it follows that FtsN initially localizes to division sites in a SPOR-independent manner. Here, we show that the cytoplasmic and transmembrane domains of FtsN (FtsN(Cyto - TM)) facilitated localization of FtsN independently of its SPOR domain but dependent on the early cell division protein FtsA. In addition, SPOR-independent localization preceded SPOR-dependent localization, providing a mechanism for the initial localization of FtsN. In support of the role of FtsNCyto - TM in FtsN function, a variant of FtsN lacking the cytoplasmic domain localized to the divisome but failed to complement an ftsN deletion unless it was overproduced. Simultaneous removal of the cytoplasmic and SPOR domains abolished localization and complementation. These data support a model in which FtsA-FtsN interaction recruits FtsN to the divisome, where it can then stimulate the peptidoglycan remodelling activities required for SPOR-dependent localization.
authors.affiliation: Department of Microbiology and Molecular Genetics, University of Texas Medical School at Houston, Houston, TX, USA.
authors.forename: Kimberly K
authors.initials: KK
authors.lastname: Busiek
authors.forename: William
authors.initials: W
authors.lastname: Margolin
mesh_terms: D002455:Cell Division; D002473:Cell Wall / Q000378:metabolism; D004252:DNA Mutational Analysis; D004926:Escherichia coli / Q000201:enzymology* / Q000378:metabolism / Q000502:physiology*; D029968:Escherichia coli Proteins / Q000235:genetics / Q000378:metabolism*; D008565:Membrane Proteins / Q000235:genetics / Q000378:metabolism*; D010457:Peptidoglycan / Q000378:metabolism; D017434:Protein Structure, Tertiary
chemical_list: D029968:Escherichia coli Proteins; C489310:FtsA protein, E coli; C098711:FtsN protein, E coli; D008565:Membrane Proteins; D010457:Peptidoglycan
references.citation: Mol Microbiol. 2007 Dec;66(6):1396-415
references.pmid: 17986188
references.citation: J Bacteriol. 2004 Nov;186(22):7736-44
references.pmid: 15516588
references.citation: EMBO J. 2002 Feb 15;21(4):685-93
references.pmid: 11847116
references.citation: J Biol Chem. 2013 Jul 19;288(29):20830-20836
references.pmid: 23740256
references.citation: Mol Microbiol. 2001 Jan;39(1):47-53
references.pmid: 11123687
references.citation: Trends Cell Biol. 1997 Sep;7(9):362-7
references.pmid: 17708981
references.citation: Mol Microbiol. 2010 May;76(3):760-71
references.pmid: 20345660
references.citation: EMBO J. 1999 May 4;18(9):2372-83
references.pmid: 10228152
references.citation: J Bacteriol. 2007 Jan;189(2):633-45
references.pmid: 16980443
references.citation: Curr Biol. 2005 Jul 12;15(13):R514-26
references.pmid: 16005287
references.citation: Mol Microbiol. 2005 Mar;55(6):1631-45
references.pmid: 15752189
references.citation: Mol Microbiol. 2012 Jan;83(1):151-67
references.pmid: 22111832
references.citation: J Bacteriol. 1996 Mar;178(5):1328-34
references.pmid: 8631709
references.citation: J Bacteriol. 2005 Jan;187(1):320-8
references.pmid: 15601716
references.citation: Mol Microbiol. 2014 Apr;92(1):1-9
references.pmid: 24506818
references.citation: J Bacteriol. 2009 Dec;191(24):7381-2
references.pmid: 19854895
references.citation: Proc Natl Acad Sci U S A. 2009 Dec 22;106(51):21872-7
references.pmid: 19995973
references.citation: Proc Natl Acad Sci U S A. 2003 Apr 1;100(7):4197-202
references.pmid: 12634424
references.citation: J Bacteriol. 2002 Oct;184(20):5572-82
references.pmid: 12270814
references.citation: J Bacteriol. 1997 Aug;179(16):5094-103
references.pmid: 9260951
references.citation: Nature. 1991 Nov 14;354(6349):161-4
references.pmid: 1944597
references.citation: Mol Microbiol. 2005 Mar;55(6):1722-34
references.pmid: 15752196
references.citation: J Bacteriol. 1999 Jan;181(2):508-20
references.pmid: 9882665
references.citation: J Biol Chem. 2007 Dec 14;282(50):36394-402
references.pmid: 17938168
references.citation: EMBO J. 2000 Jun 1;19(11):2719-27
references.pmid: 10835369
references.citation: J Bacteriol. 2012 Apr;194(8):1989-2000
references.pmid: 22328664
references.citation: J Bacteriol. 1993 Jun;175(12):3790-7
references.pmid: 8509333
references.citation: Genes Dev. 2005 Jan 1;19(1):127-37
references.pmid: 15630023
references.citation: Mol Microbiol. 2008 Feb;67(3):558-69
references.pmid: 18186792
references.citation: J Bacteriol. 2013 Jul;195(13):3022-34
references.pmid: 23625845
references.citation: J Bacteriol. 2009 Dec;191(24):7383-401
references.pmid: 19684127
references.citation: Proc Natl Acad Sci U S A. 1996 Nov 12;93(23):12998-3003
references.pmid: 8917533
references.citation: Mol Microbiol. 1997 Jul;25(2):303-9
references.pmid: 9282742
references.citation: J Bacteriol. 2004 Oct;186(20):6728-37
references.pmid: 15466024
references.citation: Mol Microbiol. 2005 Oct;58(2):596-612
references.pmid: 16194242
references.citation: J Bacteriol. 2010 Jan;192(1):242-55
references.pmid: 19880599
references.citation: J Bacteriol. 2011 Sep;193(18):4973-83
references.pmid: 21764913
references.citation: Mol Microbiol. 2007 Jun;64(5):1289-305
references.pmid: 17542921
references.citation: J Bacteriol. 2005 Apr;187(7):2233-43
references.pmid: 15774864
references.citation: Mol Microbiol. 2006 Jul;61(1):5-8
references.pmid: 16824090
references.citation: J Bacteriol. 1995 Jul;177(14):4121-30
references.pmid: 7608087
references.citation: Mol Microbiol. 2010 Jul;77(2):384-98
references.pmid: 20497333