title: Identification of key structural elements for neuronal calcium sensor-1 function in the regulation of the temperature-dependency of locomotion in C. elegans.
abstract: BACKGROUND
  Intracellular Ca2+ regulates many aspects of neuronal function through Ca2+ binding to EF hand-containing Ca2+ sensors that in turn bind target proteins to regulate their function. Amongst the sensors are the neuronal calcium sensor (NCS) family of proteins that are involved in multiple neuronal signalling pathways. Each NCS protein has specific and overlapping targets and physiological functions and specificity is likely to be determined by structural features within the proteins. Common to the NCS proteins is the exposure of a hydrophobic groove, allowing target binding in the Ca2+-loaded form. Structural analysis of NCS protein complexes with target peptides has indicated common and distinct aspects of target protein interaction. Two key differences between NCS proteins are the size of the hydrophobic groove that is exposed for interaction and the role of their non-conserved C-terminal tails.
  
  
  RESULTS
  We characterised the role of NCS-1 in a temperature-dependent locomotion assay in C. elegans and identified a distinct phenotype in the ncs-1 null in which the worms do not show reduced locomotion at actually elevated temperature. Using rescue of this phenotype we showed that NCS-1 functions in AIY neurons. Structure/function analysis introducing single or double mutations within the hydrophobic groove based on information from characterised target complexes established that both N- and C-terminal pockets of the groove are functionally important and that deletion of the C-terminal tail of NCS-1 did not impair its ability to rescue.
  
  
  CONCLUSIONS
  The current work has allowed physiological assessment of suggestions from structural studies on the key structural features that underlie the interaction of NCS-1 with its target proteins. The results are consistent with the notion that full length of the hydrophobic groove is required for the regulatory interactions underlying NCS-1 function whereas the C-terminal tail of NCS-1 is not essential. This has allowed discrimination between two potential modes of interaction of NCS-1 with its targets.
authors: Department of Cellular and Molecular Physiology, The Physiological Laboratory, Institute of Translational Medicine, University of Liverpool, Liverpool, L69 3BX, UK. burgoyne@liverpool.ac.uk.
authors: Victoria M
authors: VM
authors: Martin
authors: James R
authors: JR
authors: Johnson
authors: Lee P
authors: LP
authors: Haynes
authors: Jeff W
authors: JW
authors: Barclay
authors: Robert D
authors: RD
authors: Burgoyne
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D017173:Caenorhabditis elegans / Q000502:physiology*; D029742:Caenorhabditis elegans Proteins / Q000737:chemistry* / Q000378:metabolism*; D002135:Calcium-Binding Proteins / Q000737:chemistry* / Q000378:metabolism*; D006801:Humans; D057927:Hydrophobic and Hydrophilic Interactions; D008124:Locomotion / Q000502:physiology*; D008958:Models, Molecular; D008969:Molecular Sequence Data; D009154:Mutation / Q000235:genetics; D009419:Nerve Tissue Proteins / Q000737:chemistry* / Q000378:metabolism*; D051580:Neuronal Calcium-Sensor Proteins / Q000737:chemistry* / Q000378:metabolism*; D009474:Neurons / Q000378:metabolism; D009479:Neuropeptides / Q000737:chemistry* / Q000378:metabolism*; D017384:Sequence Deletion; D013329:Structure-Activity Relationship; D013696:Temperature
chemical_list: D029742:Caenorhabditis elegans Proteins; D002135:Calcium-Binding Proteins; C000589452:NCS-1 protein, C elegans; D009419:Nerve Tissue Proteins; D051580:Neuronal Calcium-Sensor Proteins; D009479:Neuropeptides; C101464:frequenin calcium sensor proteins
references: Hum Mol Genet. 2008 Dec 15;17(24):3965-74
references: 18801879
references: J Neurosci. 2003 Jan 15;23(2):373-6
references: 12533596
references: J Biol Chem. 2004 Apr 2;279(14):14347-54
references: 14726528
references: Proc Natl Acad Sci U S A. 1999 Aug 31;96(18):10477-82
references: 10468634
references: J Biol Chem. 2000 Dec 22;275(51):40082-7
references: 11006299
references: J Biol Chem. 2001 Apr 13;276(15):11949-55
references: 11092894
references: Neuron. 2001 Apr;30(1):241-8
references: 11343658
references: J Neurosci. 2012 May 16;32(20):7042-51
references: 22593072
references: J Biol Chem. 2004 Mar 26;279(13):12744-54
references: 14722091
references: Nat Cell Biol. 1999 Aug;1(4):234-41
references: 10559922
references: Biochim Biophys Acta. 2012 Aug;1820(8):1205-13
references: 22020049
references: J Neurosci. 2011 Aug 10;31(32):11718-27
references: 21832201
references: Neuron. 1993 Jul;11(1):15-28
references: 8101711
references: Nat Neurosci. 2007 Jan;10(1):32-9
references: 17187064
references: J Biol Chem. 2009 Feb 20;284(8):4960-7
references: 19109250
references: Biochem J. 2001 Jan 1;353(Pt 1):1-12
references: 11115393
references: J Mol Biol. 2012 Mar 16;417(1-2):51-64
references: 22227393
references: Dev Cell. 2004 Jun;6(6):757-70
references: 15177025
references: J Physiol. 2006 Apr 1;572(Pt 1):165-72
references: 16469785
references: J Neurobiol. 2003 Aug;56(2):178-97
references: 12838583
references: J Biol Chem. 2006 Jun 30;281(26):18098-111
references: 16638749
references: Mol Brain. 2012 Jan 23;5(1):2
references: 22269068
references: Nat Rev Neurosci. 2007 Mar;8(3):182-93
references: 17311005
references: PLoS One. 2010 May 07;5(5):e10534
references: 20479890
references: Neurosci Lett. 1997 Apr 4;225(2):126-8
references: 9147390
references: Proteomics. 2006 Mar;6(6):1822-32
references: 16470652
references: Hum Mol Genet. 2003 Jun 15;12(12):1415-25
references: 12783849
references: Biochemistry. 2008 Feb 26;47(8):2252-64
references: 18201103
references: Trends Neurosci. 2004 Apr;27(4):203-9
references: 15046879
references: J Biol Chem. 2006 Dec 1;281(48):37237-45
references: 17020884
references: Neuron. 1997 Aug;19(2):345-57
references: 9292724
references: WormBook. 2013 Aug 13;:1-106
references: 24081909
references: J Biol Chem. 2011 Apr 8;286(14):12565-77
references: 21288895
references: J Biol Chem. 2002 Apr 19;277(16):14227-37
references: 11836243
references: Mol Biol Cell. 2009 Jan;20(1):43-55
references: 18923141
references: Traffic. 2007 Aug;8(8):1080-92
references: 17555535
references: Neuron. 2008 Dec 26;60(6):1095-111
references: 19109914
references: Biochem J. 2009 Feb 15;418(1):73-80
references: 19032153
references: J Neurosci. 2002 Oct 1;22(19):8476-86
references: 12351722
references: J Biol Chem. 2005 Feb 18;280(7):6047-54
references: 15576365
references: Development. 2001 Jun;128(11):1951-69
references: 11493519
references: EMBO Rep. 2011 Jul 08;12(8):855-62
references: 21738224
references: Neuron. 2004 Feb 19;41(4):573-86
references: 14980206
references: EMBO J. 2006 Jan 25;25(2):312-22
references: 16407969
references: J Cell Sci. 2009 Nov 15;122(Pt 22):4109-21
references: 19861494
references: Science. 2002 Mar 22;295(5563):2276-9
references: 11910115
references: Genetics. 1974 May;77(1):71-94
references: 4366476
references: Bioinformatics. 2006 Jan 15;22(2):195-201
references: 16301204
references: Nat Neurosci. 2003 Oct;6(10):1031-8
references: 12947410
references: Biochem J. 2011 Apr 15;435(2):441-50
references: 21299498
references: BMC Biol. 2012 Oct 31;10:85
references: 23114012
references: Nature. 1997 Sep 11;389(6647):198-202
references: 9296500
references: Neuron. 2004 Feb 19;41(4):587-98
references: 14980207
references: Neuron. 2009 Sep 10;63(5):643-56
references: 19755107
references: J Clin Invest. 2006 Jun;116(6):1668-74
references: 16691292
references: Curr Opin Neurobiol. 2007 Dec;17(6):712-9
references: 18242074
references: J Biol Chem. 2007 Oct 19;282(42):30949-59
references: 17720810
references: Trends Neurosci. 1996 Dec;19(12):547-54
references: 8961484
references: J Cell Biol. 2006 Mar 27;172(7):1081-91
references: 16549499
references: Cold Spring Harb Perspect Biol. 2010 Aug;2(8):a004085
references: 20668007
references: Curr Opin Neurobiol. 2013 Feb;23(1):92-9
references: 23063296
references: Genesis. 2000 Apr;26(4):234-9
references: 10748460
references: PLoS One. 2011;6(11):e27779
references: 22114693
references: Nat Struct Mol Biol. 2006 Nov;13(11):987-95
references: 17057713
references: J Cell Biol. 2003 Nov 24;163(4):715-21
references: 14638856
references: J Biol Chem. 1998 Aug 28;273(35):22768-72
references: 9712909