title: Bacterial cytological profiling rapidly identifies the cellular pathways targeted by antibacterial molecules.
abstract: Identifying the mechanism of action for antibacterial compounds is essential for understanding how bacteria interact with one another and with other cell types and for antibiotic discovery efforts, but determining a compound's mechanism of action remains a serious challenge that limits both basic research and antibacterial discovery programs. Here, we show that bacterial cytological profiling (BCP) is a rapid and powerful approach for identifying the cellular pathway affected by antibacterial molecules. BCP can distinguish between inhibitors that affect different cellular pathways as well as different targets within the same pathway. We use BCP to demonstrate that spirohexenolide A, a spirotetronate that is active against methicillin-resistant Staphylococcus aureus, rapidly collapses the proton motive force. BCP offers a simple, one-step assay that can be broadly applied, solving the longstanding problem of how to rapidly determine the cellular target of thousands of compounds.
authors.affiliation: Division of Biological Sciences, University of California, San Diego, CA 92093.
authors.forename: Poochit
authors.initials: P
authors.lastname: Nonejuie
authors.forename: Michael
authors.initials: M
authors.lastname: Burkart
authors.forename: Kit
authors.initials: K
authors.lastname: Pogliano
authors.forename: Joe
authors.initials: J
authors.lastname: Pogliano
mesh_terms: D000900:Anti-Bacterial Agents / Q000494:pharmacology*; D003584:Cytological Techniques / Q000379:methods*; D004353:Drug Evaluation, Preclinical / Q000379:methods*; D024901:Drug Resistance, Multiple, Bacterial / Q000235:genetics*; D005434:Flow Cytometry; D006576:Heterocyclic Compounds, 4 or More Rings / Q000494:pharmacology; D008826:Microbial Sensitivity Tests; D008856:Microscopy, Fluorescence; D025341:Principal Component Analysis; D018892:Proton-Motive Force / Q000187:drug effects; D013211:Staphylococcus aureus / Q000187:drug effects*
chemical_list: D000900:Anti-Bacterial Agents; D006576:Heterocyclic Compounds, 4 or More Rings; C547347:spirohexenolide A
keywords: antibiotic resistance; drug screening; high throughput; pharmacology; susceptibility
references.citation: Clin Microbiol Rev. 2011 Jan;24(1):71-109
references.pmid: 21233508
references.citation: Science. 1999 Oct 29;286(5441):971-4
references.pmid: 10542155
references.citation: Proc Natl Acad Sci U S A. 2001 Dec 18;98(26):14925-30
references.pmid: 11734630
references.citation: Nat Rev Drug Discov. 2007 Jan;6(1):29-40
references.pmid: 17159923
references.citation: Appl Environ Microbiol. 1992 Jul;58(7):2255-9
references.pmid: 1637163
references.citation: Chem Biol. 2009 Aug 28;16(8):826-36
references.pmid: 19716473
references.citation: J Struct Biol. 2006 Nov;156(2):284-91
references.pmid: 16934488
references.citation: Appl Microbiol Biotechnol. 2006 Mar;70(2):140-50
references.pmid: 16391922
references.citation: Biochemistry. 1974 Jul 30;13(16):3315-30
references.pmid: 4842277
references.citation: Science. 2012 Jun 8;336(6086):1268-73
references.pmid: 22674334
references.citation: J Mol Biol. 2004 Nov 5;343(5):1183-94
references.pmid: 15491605
references.citation: J Org Chem. 2009 Dec 4;74(23):9054-61
references.pmid: 19883063
references.citation: J Bacteriol. 2009 Jul;191(13):4180-5
references.pmid: 19395497
references.citation: Mol Microbiol. 2012 May;84(3):486-500
references.pmid: 22469514
references.citation: Antimicrob Agents Chemother. 2011 Jan;55(1):311-20
references.pmid: 20937782
references.citation: Curr Protoc Pharmacol. 2009 Dec;Chapter 13:Unit 13A.7
references.pmid: 22294390
references.citation: J Nat Prod. 2013 May 24;76(5):817-23
references.pmid: 23659282
references.citation: Annu Rev Biochem. 2009;78:119-46
references.pmid: 19231985
references.citation: Chem Rev. 2011 Sep 14;111(9):5492-505
references.pmid: 21786783
references.citation: Annu Rev Immunol. 2010;28:623-67
references.pmid: 20192812
references.citation: J Biol Chem. 2001 Jan 19;276(3):1772-9
references.pmid: 11038353
references.citation: Chembiochem. 2005 Jan;6(1):33-9
references.pmid: 15568196
references.citation: Cell. 2011 Jan 7;144(1):143-56
references.pmid: 21185072
references.citation: Nature. 2012 Sep 13;489(7415):242-9
references.pmid: 22972297
references.citation: Prog Drug Res. 2007;64:79, 81-101
references.pmid: 17195472
references.citation: Cell. 2005 Apr 22;121(2):307-17
references.pmid: 15851036
references.citation: Cell. 2006 Feb 24;124(4):837-48
references.pmid: 16497592
references.citation: Antimicrob Agents Chemother. 2005 Feb;49(2):749-59
references.pmid: 15673760
references.citation: Mol Microbiol. 1999 Feb;31(4):1149-59
references.pmid: 10096082
references.citation: Chem Biol. 2004 Oct;11(10):1423-30
references.pmid: 15489169
references.citation: Antimicrob Agents Chemother. 1992 Oct;36(10):2118-24
references.pmid: 1280055
references.citation: Nature. 2002 Jan 24;415(6870):389-95
references.pmid: 11807545
references.citation: Curr Opin Drug Discov Devel. 2007 Mar;10(2):160-6
references.pmid: 17436551
references.citation: Biochem Biophys Res Commun. 2006 Nov 3;349(4):1250-7
references.pmid: 16978582