title: Single-cell E. coli response to an instantaneously applied chemotactic signal.
abstract: In response to an attractant or repellant, an Escherichia coli cell controls the rotational direction of its flagellar motor by a chemotaxis system. When an E. coli cell senses an attractant, a reduction in the intracellular concentration of a chemotaxis protein, phosphorylated CheY (CheY-P), induces counterclockwise (CCW) rotation of the flagellar motor, and this cellular response is thought to occur in several hundred milliseconds. Here, to measure the signaling process occurring inside a single E. coli cell, including the recognition of an attractant by a receptor cluster, the inactivation of histidine kinase CheA, and the diffusion of CheY and CheY-P molecules, we applied a serine stimulus by instantaneous photorelease from a caged compound and examined the cellular response at a temporal resolution of several hundred microseconds. We quantified the clockwise (CW) and CCW durations immediately after the photorelease of serine as the response time and the duration of the response, respectively. The results showed that the response time depended on the distance between the receptor and motor, indicating that the decreased CheY-P concentration induced by serine propagates through the cytoplasm from the receptor-kinase cluster toward the motor with a timing that is explained by the diffusion of CheY and CheY-P molecules. The response time included 240 ms for enzymatic reactions in addition to the time required for diffusion of the signaling molecule. The measured response time and duration of the response also revealed that the E. coli cell senses a similar serine concentration regardless of whether the serine concentration is increasing or decreasing. These detailed quantitative findings increase our understanding of the signal transduction process that occurs inside cells during bacterial chemotaxis.
authors.affiliation: Graduate School of Life Sciences, Tohoku University, Sendai, Japan.
authors.forename: Takashi
authors.initials: T
authors.lastname: Sagawa
authors.affiliation: Graduate School of Life Sciences, Tohoku University, Sendai, Japan.
authors.forename: Yu
authors.initials: Y
authors.lastname: Kikuchi
authors.affiliation: Graduate School of Life Sciences, Tohoku University, Sendai, Japan; Institute of Multidisciplinary Research for Advanced Materials, Tohoku University, Sendai, Japan.
authors.forename: Yuichi
authors.initials: Y
authors.lastname: Inoue
authors.affiliation: Institute of Multidisciplinary Research for Advanced Materials, Tohoku University, Sendai, Japan.
authors.forename: Hiroto
authors.initials: H
authors.lastname: Takahashi
authors.affiliation: Institute of Multidisciplinary Research for Advanced Materials, Tohoku University, Sendai, Japan.
authors.forename: Takahiro
authors.initials: T
authors.lastname: Muraoka
authors.affiliation: Institute of Multidisciplinary Research for Advanced Materials, Tohoku University, Sendai, Japan.
authors.forename: Kazushi
authors.initials: K
authors.lastname: Kinbara
authors.affiliation: Graduate School of Life Sciences, Tohoku University, Sendai, Japan; Institute of Multidisciplinary Research for Advanced Materials, Tohoku University, Sendai, Japan.
authors.forename: Akihiko
authors.initials: A
authors.lastname: Ishijima
authors.affiliation: Graduate School of Life Sciences, Tohoku University, Sendai, Japan; Institute of Multidisciplinary Research for Advanced Materials, Tohoku University, Sendai, Japan. Electronic address: f-hajime@tagen.tohoku.ac.jp.
authors.forename: Hajime
authors.initials: H
authors.lastname: Fukuoka
mesh_terms: D001426:Bacterial Proteins / Q000378:metabolism*; D002633:Chemotaxis; D004926:Escherichia coli / Q000187:drug effects / Q000378:metabolism* / Q000502:physiology; D029968:Escherichia coli Proteins; D000071677:Histidine Kinase; D008565:Membrane Proteins / Q000378:metabolism*; D000072236:Methyl-Accepting Chemotaxis Proteins; D011930:Reaction Time; D012694:Serine / Q000494:pharmacology; D059010:Single-Cell Analysis
chemical_list: D001426:Bacterial Proteins; D029968:Escherichia coli Proteins; D008565:Membrane Proteins; D000072236:Methyl-Accepting Chemotaxis Proteins; C063010:MotA protein, Bacteria; C064779:MotB protein, Bacteria; C000607606:cheY protein, E coli; D012694:Serine; D000071677:Histidine Kinase; C000607608:cheA protein, E coli
references.citation: J Bacteriol. 2007 Mar;189(5):1756-64
references.pmid: 17189361
references.citation: Science. 2000 Mar 3;287(5458):1652-5
references.pmid: 10698740
references.citation: Proc Natl Acad Sci U S A. 2011 Aug 2;108(31):E365-73
references.pmid: 21730169
references.citation: Biophys J. 1999 Mar;76(3):1706-19
references.pmid: 10049350
references.citation: Elife. 2014 Feb 11;3:e01916
references.pmid: 24520165
references.citation: J Bacteriol. 2014 Feb;196(3):641-9
references.pmid: 24272777
references.citation: J Biol Chem. 1986 Aug 15;261(23):10814-20
references.pmid: 3015942
references.citation: Proc Natl Acad Sci U S A. 1995 Oct 10;92(21):9757-61
references.pmid: 7568212
references.citation: Proc Natl Acad Sci U S A. 2009 Oct 6;106(40):17181-6
references.pmid: 19805102
references.citation: Science. 2010 Feb 5;327(5966):685-9
references.pmid: 20133571
references.citation: J Bacteriol. 2004 Jun;186(12):3687-94
references.pmid: 15175281
references.citation: Biophys J. 2011 May 4;100(9):2193-200
references.pmid: 21539787
references.citation: J Bacteriol. 1985 Jan;161(1):51-9
references.pmid: 3881399
references.citation: J Bacteriol. 2003 Apr;185(7):2354-61
references.pmid: 12644507
references.citation: Nature. 1982 Apr 29;296(5860):855-7
references.pmid: 7040985
references.citation: Nature. 2000 Jan 27;403(6768):444-7
references.pmid: 10667798
references.citation: PLoS Comput Biol. 2006 Apr;2(4):e39
references.pmid: 16683020
references.citation: Proc Natl Acad Sci U S A. 1998 Jan 6;95(1):201-6
references.pmid: 9419353
references.citation: J Mol Biol. 2011 Aug 19;411(3):554-66
references.pmid: 21718703
references.citation: Mol Syst Biol. 2009;5:238
references.pmid: 19156130
references.citation: Biophys J. 1993 Dec;65(6):2368-82
references.pmid: 8312476
references.citation: Sci Signal. 2014 Apr 01;7(319):ra32
references.pmid: 24692593
references.citation: Proc Natl Acad Sci U S A. 1993 Oct 1;90(19):8787-91
references.pmid: 8415608
references.citation: Proc Natl Acad Sci U S A. 2002 Sep 3;99(18):11611-5
references.pmid: 12186970
references.citation: J Mol Biol. 1998 May 8;278(3):507-14
references.pmid: 9600834
references.citation: Nat Rev Mol Cell Biol. 2004 Dec;5(12):1024-37
references.pmid: 15573139
references.citation: J Bacteriol. 2005 Jan;187(1):45-53
references.pmid: 15601687
references.citation: Proc Natl Acad Sci U S A. 2004 Dec 7;101(49):17072-7
references.pmid: 15569922
references.citation: Cell. 2000 Feb 4;100(3):357-65
references.pmid: 10676817
references.citation: Proc Natl Acad Sci U S A. 2002 Oct 1;99(20):12669-74
references.pmid: 12232047
references.citation: Nature. 1998 May 7;393(6680):85-8
references.pmid: 9590695
references.citation: J Biol Chem. 2011 Dec 9;286(49):42200-42210
references.pmid: 21979954
references.citation: Biochemistry. 1997 Feb 25;36(8):2030-40
references.pmid: 9047301