title: Sodium solute symporter and cadherin proteins act as Bacillus thuringiensis Cry3Ba toxin functional receptors in Tribolium castaneum.
abstract: Understanding how Bacillus thuringiensis (Bt) toxins interact with proteins in the midgut of susceptible coleopteran insects is crucial to fully explain the molecular bases of Bt specificity and insecticidal action. In this work, aminopeptidase N (TcAPN-I), E-cadherin (TcCad1), and sodium solute symporter (TcSSS) have been identified by ligand blot as putative Cry3Ba toxin-binding proteins in Tribolium castaneum (Tc) larvae. RNA interference knockdown of TcCad1 or TcSSS proteins resulted in decreased susceptibility to Cry3Ba toxin, demonstrating the Cry toxin receptor functionality for these proteins. In contrast, TcAPN-I silencing had no effect on Cry3Ba larval toxicity, suggesting that this protein is not relevant in the Cry3Ba toxin mode of action in Tc. Remarkable features of TcSSS protein were the presence of cadherin repeats in its amino acid sequence and that a TcSSS peptide fragment containing a sequence homologous to a binding epitope found in Manduca sexta and Tenebrio molitor Bt cadherin functional receptors enhanced Cry3Ba toxicity. This is the first time that the involvement of a sodium solute symporter protein as a Bt functional receptor has been demonstrated. The role of this novel receptor in Bt toxicity against coleopteran insects together with the lack of receptor functionality of aminopeptidase N proteins might account for some of the differences in toxin specificity between Lepidoptera and Coleoptera insect orders.
authors: Departamento de Genética, Facultad de Ciencias Biológicas, Universidad de Valencia, Burjassot 46100, Valencia, Spain.
authors: Estefanía
authors: E
authors: Contreras
authors: Michael
authors: M
authors: Schoppmeier
authors: M Dolores
authors: MD
authors: Real
authors: Carolina
authors: C
authors: Rausell
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D000083722:Bacillus thuringiensis Toxins; D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism*; D001665:Binding Sites / Q000235:genetics; D018826:CD13 Antigens / Q000235:genetics / Q000378:metabolism; D015820:Cadherins / Q000235:genetics / Q000378:metabolism; D004731:Endotoxins / Q000235:genetics / Q000378:metabolism*; D000939:Epitopes / Q000235:genetics / Q000378:metabolism; D006460:Hemolysin Proteins / Q000235:genetics / Q000378:metabolism*; D015151:Immunoblotting; D019476:Insect Proteins / Q000235:genetics / Q000378:metabolism*; D008969:Molecular Sequence Data; D011485:Protein Binding; D034622:RNA Interference; D011956:Receptors, Cell Surface / Q000235:genetics / Q000378:metabolism; D017386:Sequence Homology, Amino Acid; D012964:Sodium / Q000378:metabolism; D027981:Symporters / Q000235:genetics / Q000378:metabolism; D014231:Tribolium / Q000235:genetics / Q000378:metabolism*
chemical_list: D000083722:Bacillus thuringiensis Toxins; D001426:Bacterial Proteins; D015820:Cadherins; D004731:Endotoxins; D000939:Epitopes; D006460:Hemolysin Proteins; D019476:Insect Proteins; D011956:Receptors, Cell Surface; D027981:Symporters; C030956:insecticidal crystal protein, Bacillus Thuringiensis; D012964:Sodium; D018826:CD13 Antigens
keywords: Cadherins; Host-Pathogen Interactions; RNA Interference (RNAi); Receptors; Transporters
references: Appl Environ Microbiol. 2011 Jan;77(1):24-31
references: 21037295
references: Biochem J. 2009 Nov 11;424(2):191-200
references: 19732034
references: J Biol Chem. 1995 Mar 10;270(10):5490-4
references: 7890666
references: Trends Microbiol. 2005 Aug;13(8):347-50
references: 15967665
references: Insect Biochem Mol Biol. 2002 Sep;32(9):1025-36
references: 12213239
references: J Proteome Res. 2009 Aug;8(8):3889-98
references: 19545177
references: Pest Manag Sci. 2011 Sep;67(9):1076-81
references: 21495115
references: J Biol Chem. 2004 Jul 2;279(27):28051-6
references: 15123702
references: FEBS J. 2013 Apr;280(8):1782-94
references: 23432933
references: Microbiol Mol Biol Rev. 2007 Jun;71(2):255-81
references: 17554045
references: Proc Natl Acad Sci U S A. 2007 Aug 28;104(35):13901-6
references: 17724346
references: J Biol Chem. 2001 Aug 3;276(31):28906-12
references: 11384982
references: PLoS Genet. 2010 Dec 16;6(12):e1001248
references: 21187898
references: J Mol Microbiol Biotechnol. 2012;22(2):105-13
references: 22627175
references: Insect Biochem Mol Biol. 2009 Apr;39(4):279-86
references: 19272330
references: Insect Biochem Mol Biol. 2010 Feb;40(2):138-45
references: 20079436
references: Insect Biochem Mol Biol. 2007 Mar;37(3):189-201
references: 17296494
references: Insect Biochem Mol Biol. 2009 Nov;39(11):824-32
references: 19815067
references: J Biol Chem. 2009 Jul 3;284(27):18401-10
references: 19416969
references: Biochem Biophys Res Commun. 1991 Dec 31;181(3):1089-93
references: 1662492
references: Biochim Biophys Acta. 2004 Jan 28;1660(1-2):99-105
references: 14757225
references: Arch Insect Biochem Physiol. 2010 Feb;73(2):61-73
references: 19847794
references: PLoS One. 2013;8(1):e55330
references: 23372850
references: Eur J Biochem. 2004 Aug;271(15):3127-35
references: 15265032
references: Nat Methods. 2011 Sep 29;8(10):785-6
references: 21959131
references: Mol Biol Evol. 1987 Jul;4(4):406-25
references: 3447015
references: Dev Comp Immunol. 2012 May;37(1):184-92
references: 21925538
references: Mol Biol Evol. 2011 Oct;28(10):2731-9
references: 21546353
references: J Biol Chem. 2002 Dec 6;277(49):46849-51
references: 12377776
references: Mol Microbiol. 1994 Feb;11(3):429-36
references: 7908713
references: Protein Sci. 1995 Aug;4(8):1587-95
references: 8520485
references: Biochem Biophys Res Commun. 2007 Oct 19;362(2):437-42
references: 17714689
references: J Microbiol Methods. 2010 Feb;80(2):219-21
references: 20026205
references: Science. 2007 Dec 7;318(5856):1640-2
references: 17975031
references: Nature. 1966 Aug 27;211(5052):969-70
references: 5968307
references: BMC Genomics. 2005 Jun 24;6:96
references: 15978131
references: Insect Biochem Mol Biol. 2003 Oct;33(10):999-1010
references: 14505693
references: Biochem J. 1981 Jul 15;198(1):101-6
references: 6798972
references: Nucleic Acids Res. 2009 Jan;37(Database issue):D274-8
references: 19022853
references: Biochim Biophys Acta. 2004 Nov 17;1667(1):38-46
references: 15533304
references: FEBS Lett. 2002 Oct 2;529(1):73-7
references: 12354616
references: Proc Natl Acad Sci U S A. 2012 Jun 19;109(25):E1591-8
references: 22635270
references: Microbiol Mol Biol Rev. 2000 Jun;64(2):354-411
references: 10839820