title: A distance-weighted interaction map reveals a previously uncharacterized layer of the Bacillus subtilis spore coat.
abstract: Bacillus subtilis spores are encased in a protein assembly called the spore coat that is made up of at least 70 different proteins. Conventional electron microscopy shows the coat to be organized into two distinct layers. Because the coat is about as wide as the theoretical limit of light microscopy, quantitatively measuring the localization of individual coat proteins within the coat is challenging. We used fusions of coat proteins to green fluorescent protein to map genetic dependencies for coat assembly and to define three independent subnetworks of coat proteins. To complement the genetic data, we measured coat protein localization at subpixel resolution and integrated these two data sets to produce a distance-weighted genetic interaction map. Using these data, we predict that the coat comprises at least four spatially distinct layers, including a previously uncharacterized glycoprotein outermost layer that we name the spore crust. We found that crust assembly depends on proteins we predicted to localize to the crust. The crust may be conserved in all Bacillus spores and may play critical functions in the environment.
authors.affiliation: Center for Genomics and Systems Biology, Department of Biology, New York University, New York, NY 10003, USA.
authors.forename: Peter T
authors.initials: PT
authors.lastname: McKenney
authors.forename: Adam
authors.initials: A
authors.lastname: Driks
authors.forename: Haig A
authors.initials: HA
authors.lastname: Eskandarian
authors.forename: Paul
authors.initials: P
authors.lastname: Grabowski
authors.forename: Jonathan
authors.initials: J
authors.lastname: Guberman
authors.forename: Katherine H
authors.initials: KH
authors.lastname: Wang
authors.forename: Zemer
authors.initials: Z
authors.lastname: Gitai
authors.forename: Patrick
authors.initials: P
authors.lastname: Eichenberger
mesh_terms: D001412:Bacillus subtilis / Q000737:chemistry / Q000378:metabolism / Q000648:ultrastructure*; D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism; D015964:Gene Expression Regulation, Bacterial; D053263:Gene Regulatory Networks; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D013171:Spores, Bacterial / Q000737:chemistry / Q000648:ultrastructure*
chemical_list: D001426:Bacterial Proteins; D011993:Recombinant Fusion Proteins
references.citation: Science. 2009 Mar 6;323(5919):1354-7
references.pmid: 19265022
references.citation: J Bacteriol. 1995 Oct;177(20):5906-11
references.pmid: 7592342
references.citation: Curr Opin Microbiol. 2009 Jun;12(3):341-6
references.pmid: 19356974
references.citation: Genes Dev. 1988 Aug;2(8):1047-54
references.pmid: 3139490
references.citation: Annu Rev Microbiol. 2007;61:555-88
references.pmid: 18035610
references.citation: Nature. 2007 Aug 23;448(7156):883-8
references.pmid: 17713526
references.citation: J Bacteriol. 2005 Jul;187(13):4592-7
references.pmid: 15968070
references.citation: Bacteriol Rev. 1976 Jun;40(2):360-402
references.pmid: 786255
references.citation: PLoS Comput Biol. 2008 Nov;4(11):e1000233
references.pmid: 19043544
references.citation: Microbiology (Reading). 2004 Feb;150(Pt 2):355-363
references.pmid: 14766913
references.citation: J Bacteriol. 2001 May;183(10):3041-9
references.pmid: 11325931
references.citation: Mol Microbiol. 2006 Jan;59(2):487-502
references.pmid: 16390444
references.citation: J Cell Biol. 1963 Mar;16:579-92
references.pmid: 13999018
references.citation: Biophys J. 2008 Jan 1;94(1):251-64
references.pmid: 17766333
references.citation: J Bacteriol. 2009 May;191(10):3212-9
references.pmid: 19304857
references.citation: Nat Rev Microbiol. 2009 Sep;7(9):642-53
references.pmid: 19680248
references.citation: Curr Biol. 2009 Apr 28;19(8):694-9
references.pmid: 19345105
references.citation: Proc Natl Acad Sci U S A. 1999 Dec 7;96(25):14553-8
references.pmid: 10588743
references.citation: J Bacteriol. 2006 Nov;188(21):7440-8
references.pmid: 16936017
references.citation: J Bacteriol. 1993 Jun;175(12):3757-66
references.pmid: 8509331
references.citation: Mol Aspects Med. 2009 Dec;30(6):368-73
references.pmid: 19683018
references.citation: Anat Rec. 1971 Nov;171(3):347-68
references.pmid: 4108333
references.citation: Curr Opin Microbiol. 2004 Dec;7(6):579-86
references.pmid: 15556029
references.citation: Nature. 2007 Nov 29;450(7170):695-701
references.pmid: 18046406
references.citation: J Bacteriol. 2000 Apr;182(7):1828-33
references.pmid: 10714986
references.citation: J Bacteriol. 1999 Aug;181(16):4986-94
references.pmid: 10438771
references.citation: J Bacteriol. 2010 Jan;192(2):518-24
references.pmid: 19933362
references.citation: Plasmid. 1984 Jul;12(1):1-9
references.pmid: 6093169
references.citation: Mol Microbiol. 2001 Nov;42(4):1107-20
references.pmid: 11737650
references.citation: Mol Cell. 2008 Aug 8;31(3):406-14
references.pmid: 18691972
references.citation: Bacteriol Rev. 1976 Dec;40(4):908-62
references.pmid: 12736
references.citation: Nature. 2007 Dec 13;450(7172):973-82
references.pmid: 18075576
references.citation: Nat Rev Microbiol. 2008 Jun;6(6):455-65
references.pmid: 18483484
references.citation: Mol Microbiol. 2007 Jul;65(2):454-64
references.pmid: 17630974
references.citation: J Cell Biol. 2000 Feb 21;148(4):635-51
references.pmid: 10684247
references.citation: Microbiol Mol Biol Rev. 1999 Mar;63(1):1-20
references.pmid: 10066829
references.citation: Colloids Surf B Biointerfaces. 2010 Apr 1;76(2):512-8
references.pmid: 20074921
references.citation: Science. 2009 Nov 27;326(5957):1225-8
references.pmid: 19965466
references.citation: Genes Dev. 1994 Jan;8(2):234-44
references.pmid: 8299942
references.citation: Mol Microbiol. 2009 Nov;74(3):634-49
references.pmid: 19775244
references.citation: J Microbiol Methods. 2004 Jul;58(1):23-30
references.pmid: 15177900
references.citation: Mol Microbiol. 2008 May;68(4):947-58
references.pmid: 18399937