title: Calcium-release channels in paramecium. Genomic expansion, differential positioning and partial transcriptional elimination.
abstract: The release of Ca²⁺ from internal stores is a major source of signal Ca²⁺ in almost all cell types. The internal Ca²⁺ pools are activated via two main families of intracellular Ca²⁺-release channels, the ryanodine and the inositol 1,4,5-trisphosphate (InsP₃) receptors. Among multicellular organisms these channel types are ubiquitous, whereas in most unicellular eukaryotes the identification of orthologs is impaired probably due to evolutionary sequence divergence. However, the ciliated protozoan Paramecium allowed us to prognosticate six groups, with a total of 34 genes, encoding proteins with characteristics typical of InsP₃ and ryanodine receptors by BLAST search of the Paramecium database. We here report that these Ca²⁺-release channels may display all or only some of the characteristics of canonical InsP₃ and ryanodine receptors. In all cases, prediction methods indicate the presence of six trans-membrane regions in the C-terminal domains, thus corresponding to canonical InsP₃ receptors, while a sequence homologous to the InsP₃-binding domain is present only in some types. Only two types have been analyzed in detail previously. We now show, by using antibodies and eventually by green fluorescent protein labeling, that the members of all six groups localize to distinct organelles known to participate in vesicle trafficking and, thus, may provide Ca²⁺ for local membrane-membrane interactions. Whole genome duplication can explain radiation within the six groups. Comparative and evolutionary evaluation suggests derivation from a common ancestor of canonical InsP₃ and ryanodine receptors. With one group we could ascertain, to our knowledge for the first time, aberrant splicing in one thoroughly analyzed Paramecium gene. This yields truncated forms and, thus, may indicate a way to pseudogene formation. No comparable analysis is available for any other, free-living or parasitic/pathogenic protozoan.
authors.affiliation: Department of Biology, University of Konstanz, Konstanz, Germany. Eva.Ladenburger@uni-konstanz.de
authors.forename: Eva-Maria
authors.initials: EM
authors.lastname: Ladenburger
authors.forename: Helmut
authors.initials: H
authors.lastname: Plattner
mesh_terms: D017398:Alternative Splicing; D000595:Amino Acid Sequence; D002118:Calcium / Q000378:metabolism*; D020013:Calcium Signaling; D002462:Cell Membrane / Q000378:metabolism; D023281:Genomics; D053496:Inositol 1,4,5-Trisphosphate Receptors / Q000235:genetics* / Q000378:metabolism; D008969:Molecular Sequence Data; D015388:Organelles / Q000378:metabolism; D016805:Paramecium tetraurelia / Q000166:cytology / Q000235:genetics* / Q000378:metabolism*; D010802:Phylogeny; D012433:Ryanodine / Q000378:metabolism; D019837:Ryanodine Receptor Calcium Release Channel / Q000235:genetics* / Q000378:metabolism; D017386:Sequence Homology, Amino Acid
chemical_list: D053496:Inositol 1,4,5-Trisphosphate Receptors; D019837:Ryanodine Receptor Calcium Release Channel; D012433:Ryanodine; D002118:Calcium
references.citation: Curr Opin Cell Biol. 2004 Jun;16(3):293-9
references.pmid: 15145354
references.citation: Mol Biol Evol. 2006 Mar;23(3):691-700
references.pmid: 16364968
references.citation: J Gen Physiol. 1999 Aug;114(2):243-50
references.pmid: 10436000
references.citation: J Cell Sci. 2007 Jan 1;120(Pt 1):177-90
references.pmid: 17164292
references.citation: Cold Spring Harb Protoc. 2010 Jan;2010(1):pdb.prot5365
references.pmid: 20150124
references.citation: Cell Calcium. 2003 Aug;34(2):163-8
references.pmid: 12810058
references.citation: Cell Calcium. 1998 Nov-Dec;24(5-6):345-57
references.pmid: 10091004
references.citation: Protein Sci. 2004 Jul;13(7):1908-17
references.pmid: 15215532
references.citation: J Cell Sci. 2008 Jan 1;121(Pt 1):65-74
references.pmid: 18057024
references.citation: Biochemistry. 2009 Dec 29;48(51):12062-80
references.pmid: 19928968
references.citation: FEBS Lett. 2004 Sep 24;575(1-3):95-8
references.pmid: 15388340
references.citation: J Cell Sci. 2011 May 1;124(Pt 9):1411-24
references.pmid: 21486954
references.citation: Bioessays. 2011 Feb;33(2):103-10
references.pmid: 21140470
references.citation: Nature. 2008 Jan 17;451(7176):359-62
references.pmid: 18202663
references.citation: Methods Mol Biol. 1993;15:263-8
references.pmid: 21400284
references.citation: Nucleic Acids Res. 1994 Apr 11;22(7):1221-5
references.pmid: 8165136
references.citation: Eur J Cell Biol. 2000 Sep;79(9):642-52
references.pmid: 11043405
references.citation: FEBS Lett. 2010 May 17;584(10):1966-74
references.pmid: 20159015
references.citation: Mol Biol Cell. 2006 Feb;17(2):917-30
references.pmid: 16314392
references.citation: Eukaryot Cell. 2002 Jun;1(3):341-52
references.pmid: 12455983
references.citation: Nucleic Acids Res. 2007 Jan;35(Database issue):D439-44
references.pmid: 17142227
references.citation: Cell. 2007 Dec 14;131(6):1047-58
references.pmid: 18083096
references.citation: Cell Calcium. 2004 Nov;36(5):359-65
references.pmid: 15451620
references.citation: Biochim Biophys Acta. 2002 Jun 12;1590(1-3):159-66
references.pmid: 12063179
references.citation: Traffic. 2007 May;8(5):523-42
references.pmid: 17451555
references.citation: Protein Eng. 2001 Nov;14(11):867-74
references.pmid: 11742105
references.citation: Int Rev Cell Mol Biol. 2010;280:79-184
references.pmid: 20797682
references.citation: Nucleic Acids Res. 2009 Jul;37(Web Server issue):W465-8
references.pmid: 19429891
references.citation: Trends Pharmacol Sci. 1998 Sep;19(9):370-5
references.pmid: 9786025
references.citation: Mol Biol Evol. 2004 Oct;21(10):1855-70
references.pmid: 15215322
references.citation: Int Rev Cytol. 2001;201:115-208
references.pmid: 11057832
references.citation: J Cell Biol. 1991 Apr;113(1):103-12
references.pmid: 1848863
references.citation: J Biol Chem. 1990 Feb 5;265(4):2244-56
references.pmid: 2298749
references.citation: Proc Int Conf Intell Syst Mol Biol. 1998;6:175-82
references.pmid: 9783223
references.citation: Proc Natl Acad Sci U S A. 2002 Dec 24;99(26):16725-30
references.pmid: 12486242
references.citation: Pflugers Arch. 2003 Jul;446(4):447-54
references.pmid: 12764616
references.citation: Nature. 1998 Dec 10;396(6711):575-80
references.pmid: 9859992
references.citation: Mol Microbiol. 2000 Aug;37(4):773-87
references.pmid: 10972800
references.citation: Trends Biochem Sci. 2004 Apr;29(4):210-9
references.pmid: 15082315
references.citation: J Mol Biol. 1982 May 5;157(1):105-32
references.pmid: 7108955
references.citation: Brief Bioinform. 2004 Jun;5(2):150-63
references.pmid: 15260895
references.citation: J Cell Biol. 1997 Feb 10;136(3):597-607
references.pmid: 9024690
references.citation: Nature. 2006 Nov 9;444(7116):171-8
references.pmid: 17086204
references.citation: J Biol Chem. 2011 Jul 1;286(26):23378-87
references.pmid: 21550988
references.citation: Cell Calcium. 2003 Jul;34(1):87-96
references.pmid: 12767896
references.citation: Clin Exp Pharmacol Physiol. 2003 Oct;30(10):713-23
references.pmid: 14516409
references.citation: J Immunol. 2008 Oct 15;181(8):5289-95
references.pmid: 18832684
references.citation: Biochem Soc Trans. 2010 Dec;38(6):1413-6
references.pmid: 21118098
references.citation: Eur J Cell Biol. 2002 Sep;81(9):505-15
references.pmid: 12416727
references.citation: Eur J Cell Biol. 1999 Jan;78(1):67-77
references.pmid: 10082425
references.citation: Nat Rev Mol Cell Biol. 2000 Oct;1(1):11-21
references.pmid: 11413485
references.citation: J Cell Sci. 1992 Feb;101 ( Pt 2):449-61
references.pmid: 1629255
references.citation: EMBO Rep. 2007 Mar;8(3):236-40
references.pmid: 17330068
references.citation: Proc Natl Acad Sci U S A. 2009 Oct 20;106(42):17747-50
references.pmid: 19815527
references.citation: PLoS Genet. 2010 Oct 14;6(10):e1001155
references.pmid: 20976245
references.citation: J Biol Chem. 1994 Mar 25;269(12):9184-9
references.pmid: 8132655
references.citation: Bioessays. 2000 Nov;22(11):1035-42
references.pmid: 11056480
references.citation: J Biol Chem. 1996 Jul 26;271(30):18277-84
references.pmid: 8663526
references.citation: Int Rev Cytol. 2003;232:185-216
references.pmid: 14711119
references.citation: Eukaryot Cell. 2010 Feb;9(2):288-305
references.pmid: 20023070
references.citation: J Membr Biol. 1995 May;145(2):109-18
references.pmid: 7563013
references.citation: Curr Biol. 2004 Aug 10;14(15):1397-404
references.pmid: 15296759
references.citation: J Cell Sci. 2002 Oct 15;115(Pt 20):3935-46
references.pmid: 12244131
references.citation: J Cell Biol. 1995 Dec;131(6 Pt 1):1539-45
references.pmid: 8522610
references.citation: Pharmacol Rev. 1993 Mar;45(1):43-85
references.pmid: 8475169
references.citation: J Cell Sci. 1997 Mar;110 ( Pt 5):529-35
references.pmid: 9092935
references.citation: J Cell Sci. 2006 Sep 1;119(Pt 17):3705-17
references.pmid: 16912081
references.citation: Mol Cell Biol. 2009 Jul;29(13):3605-22
references.pmid: 19380481
references.citation: J Eukaryot Microbiol. 2001 Nov-Dec;48(6):640-6
references.pmid: 11831772
references.citation: Cell Calcium. 2005 Sep-Oct;38(3-4):253-60
references.pmid: 16115682
references.citation: Annu Rev Plant Biol. 2010;61:593-620
references.pmid: 20192754
references.citation: J Cell Sci. 2009 Jul 15;122(Pt 14):2337-50
references.pmid: 19571113
references.citation: J Biol Chem. 2008 Mar 14;283(11):7206-18
references.pmid: 18096697
references.citation: J Cell Biol. 1991 Jun;113(6):1295-304
references.pmid: 2045413
references.citation: Nat Rev Mol Cell Biol. 2003 Jul;4(7):517-29
references.pmid: 12838335
references.citation: Trends Biochem Sci. 2000 Feb;25(2):48-50
references.pmid: 10664581
references.citation: Q Rev Biophys. 2001 Feb;34(1):61-104
references.pmid: 11388090
references.citation: Traffic. 2006 Apr;7(4):440-55
references.pmid: 16536742
references.citation: Mol Biol Evol. 2006 Nov;23(11):2039-48
references.pmid: 16891379
references.citation: Mol Biol Evol. 2006 Aug;23(8):1613-27
references.pmid: 16751258
references.citation: Nucleic Acids Res. 1997 Sep 1;25(17):3389-402
references.pmid: 9254694
references.citation: Nature. 1989 Jun 8;339(6224):439-45
references.pmid: 2725677
references.citation: J Biol Chem. 2004 Mar 19;279(12):11967-75
references.pmid: 14707143
references.citation: Annu Rev Biochem. 2007;76:367-85
references.pmid: 17506640
references.citation: J Biol Chem. 2001 Apr 27;276(17):13509-12
references.pmid: 11278266
references.citation: J Cell Biol. 2006 Sep 25;174(7):997-1007
references.pmid: 16982801
references.citation: Proc Natl Acad Sci U S A. 2008 May 20;105(20):7177-81
references.pmid: 18477697
references.citation: Int Rev Cytol. 2000;198:277-318
references.pmid: 10804465
references.citation: J Photochem Photobiol B. 2000 Jan;54(1):35-42
references.pmid: 10739141
references.citation: Curr Biol. 2005 Aug 9;15(15):1401-6
references.pmid: 16085493
references.citation: Biol Cell. 2004 Feb;96(1):3-17
references.pmid: 15093123
references.citation: J Biol Chem. 2007 Aug 10;282(32):23096-103
references.pmid: 17526491
references.citation: Proc Natl Acad Sci U S A. 1997 Jun 24;94(13):7012-7
references.pmid: 9192683
references.citation: Mol Cell Biol. 2000 Nov;20(21):7955-70
references.pmid: 11027266
references.citation: J Biol Chem. 2008 Jan 11;283(2):751-5
references.pmid: 17999955
references.citation: Nucleic Acids Res. 1994 Nov 11;22(22):4673-80
references.pmid: 7984417
references.citation: Pharmacol Rev. 1997 Mar;49(1):53-98
references.pmid: 9085309
references.citation: J Physiol. 1991 Jun;437:461-75
references.pmid: 1716314
references.citation: Bioinformatics. 2008 Aug 1;24(15):1662-8
references.pmid: 18474507
references.citation: Methods Cell Biol. 1995;47:17-24
references.pmid: 7476483