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flp-32 Ligand/receptor silencing phenocopy faster plant pathogenic nematodes.

Atkinson LE, Stevenson M, McCoy CJ, Marks NJ, Fleming C, Zamanian M, Day TA, Kimber MJ, Maule AG, Mousley A - PLoS Pathog. (2013)

Bottom Line: This study investigates the role of flp-32 in G. pallida and shows that: (i) Gp-flp-32 encodes the peptide AMRNALVRFamide; (ii) Gp-flp-32 is expressed in the brain and ventral nerve cord of G. pallida; (iii) migration rate increases in Gp-flp-32-silenced worms; (iv) the ability of G. pallida to infect potato plant root systems is enhanced in Gp-flp-32-silenced worms; (v) a novel putative Gp-flp-32 receptor (Gp-flp-32R) is expressed in G. pallida; and, (vi) Gp-flp-32R-silenced worms also display an increase in migration rate.This work demonstrates that Gp-flp-32 plays an intrinsic role in the modulation of locomotory behaviour in G. pallida and putatively interacts with at least one novel G-protein coupled receptor (Gp-flp-32R).This is the first functional characterisation of a parasitic nematode FLP-GPCR.

View Article: PubMed Central - PubMed

Affiliation: Molecular Biosciences-Parasitology, Institute for Global Food Security, School of Biological Sciences, Queen's University Belfast, Belfast, United Kingdom.

ABSTRACT
Restrictions on nematicide usage underscore the need for novel control strategies for plant pathogenic nematodes such as Globodera pallida (potato cyst nematode) that impose a significant economic burden on plant cultivation activities. The nematode neuropeptide signalling system is an attractive resource for novel control targets as it plays a critical role in sensory and motor functions. The FMRFamide-like peptides (FLPs) form the largest and most diverse family of neuropeptides in invertebrates, and are structurally conserved across nematode species, highlighting the utility of the FLPergic system as a broad-spectrum control target. flp-32 is expressed widely across nematode species. This study investigates the role of flp-32 in G. pallida and shows that: (i) Gp-flp-32 encodes the peptide AMRNALVRFamide; (ii) Gp-flp-32 is expressed in the brain and ventral nerve cord of G. pallida; (iii) migration rate increases in Gp-flp-32-silenced worms; (iv) the ability of G. pallida to infect potato plant root systems is enhanced in Gp-flp-32-silenced worms; (v) a novel putative Gp-flp-32 receptor (Gp-flp-32R) is expressed in G. pallida; and, (vi) Gp-flp-32R-silenced worms also display an increase in migration rate. This work demonstrates that Gp-flp-32 plays an intrinsic role in the modulation of locomotory behaviour in G. pallida and putatively interacts with at least one novel G-protein coupled receptor (Gp-flp-32R). This is the first functional characterisation of a parasitic nematode FLP-GPCR.

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A novel Gp-flp-32 receptor (Gp-flp-32R) is expressed in Globodera pallida J2s and is conserved across the nematode phylum.A novel Gp-flp-32R which shows close homology to the C. elegans VRFa receptor 1 (C26F1.6) is expressed in G. pallida (A). The 389 amino acid protein has seven transmembrane helices; outside to inside helices are boxed in red; inside to outside helices are boxed in blue (A). 11 conserved residues common to rhodopsin family GPCRs are denoted by asterisks (A). Interrogation of available nematode EST, genomic and transcriptomic datasets show that at least 12 of the 16 nematode species from clades IV and V which express flp-32 also express a homologue of the Gp-flp-32R (B), demonstrating inter-clade conservation of the flp-32 receptor. B. xylophilis: Bursaphelenchus xylophilus; G. pallida: Globodera pallida; G rostochiensis: Globodera rostochiensis; M. hapla: Meloidogyne hapla; M. incognita: Meloidogyne incognita; M. paranaensis: Meloidogyne paranaensis; R. similis: Radopholus similis; S. ratti: Strongyloides ratti; A. caninum: Ancylostoma caninum; A. cantonensis: Ancylostoma cantonensis; A. ceylanicum; Ancylostoma ceylanicum; C. elegans: Caenorhabditis elegans; H. polygyrus: Heligmosomoides polygyrus; N. americanus: Necator americanus; N. brasiliensis: Nippostrongylus brasiliensis; T. circumcincta: Teladorsagia circumcincta.
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ppat-1003169-g007: A novel Gp-flp-32 receptor (Gp-flp-32R) is expressed in Globodera pallida J2s and is conserved across the nematode phylum.A novel Gp-flp-32R which shows close homology to the C. elegans VRFa receptor 1 (C26F1.6) is expressed in G. pallida (A). The 389 amino acid protein has seven transmembrane helices; outside to inside helices are boxed in red; inside to outside helices are boxed in blue (A). 11 conserved residues common to rhodopsin family GPCRs are denoted by asterisks (A). Interrogation of available nematode EST, genomic and transcriptomic datasets show that at least 12 of the 16 nematode species from clades IV and V which express flp-32 also express a homologue of the Gp-flp-32R (B), demonstrating inter-clade conservation of the flp-32 receptor. B. xylophilis: Bursaphelenchus xylophilus; G. pallida: Globodera pallida; G rostochiensis: Globodera rostochiensis; M. hapla: Meloidogyne hapla; M. incognita: Meloidogyne incognita; M. paranaensis: Meloidogyne paranaensis; R. similis: Radopholus similis; S. ratti: Strongyloides ratti; A. caninum: Ancylostoma caninum; A. cantonensis: Ancylostoma cantonensis; A. ceylanicum; Ancylostoma ceylanicum; C. elegans: Caenorhabditis elegans; H. polygyrus: Heligmosomoides polygyrus; N. americanus: Necator americanus; N. brasiliensis: Nippostrongylus brasiliensis; T. circumcincta: Teladorsagia circumcincta.

Mentions: Database mining facilitated the identification of a putative FLP-32 receptor in G. pallida, orthologous to the C. elegans VRFa receptor R1 (C26F1.6; see Fig. 7A). RACE PCR and sequencing confirmed the sequence of the putative G. pallida flp-32 receptor (Gp-flp-32R). Primers designed to confirm the open reading frame of Gp-flp-32R generated a 1,170 nucleotide cDNA sequence (GenBank accession number JQ685132), encoding a 389 aa protein (Fig. 7A). Gp-flp-32R encodes seven transmembrane helices and conserved residues at positions 48, 52, 76, 80, 136–138, 222, 273 and 318–320; the 136–138 sequence (DRF) is a common variation on the DRY motif at the cytosolic end of the third transmembrane helix and is common to rhodopsin-like GPCRs (see Fig. 7A). In a reciprocal tBLASTn search of the C. elegans non-redundant nucleotide and protein database, C. elegans C26F1.6 was returned as the top scoring hit (53% identity to Gp-flp-32R). Further interrogation of the G. pallida EST database (GenBank) and genome assembly (Wellcome Trust Sanger Institute November 2010 supercontig assembly) between August and October 2011 with C. elegans C26F1.6 did not reveal additional homologous transcripts. BLAST searches of all available nematode EST, genomic and transcriptomic resources identified 12 C26F1.6 GPCR homologues from the 16 species which express FLP-32 encoding transcripts, spanning clades IV and V (Fig. 7B).


flp-32 Ligand/receptor silencing phenocopy faster plant pathogenic nematodes.

Atkinson LE, Stevenson M, McCoy CJ, Marks NJ, Fleming C, Zamanian M, Day TA, Kimber MJ, Maule AG, Mousley A - PLoS Pathog. (2013)

A novel Gp-flp-32 receptor (Gp-flp-32R) is expressed in Globodera pallida J2s and is conserved across the nematode phylum.A novel Gp-flp-32R which shows close homology to the C. elegans VRFa receptor 1 (C26F1.6) is expressed in G. pallida (A). The 389 amino acid protein has seven transmembrane helices; outside to inside helices are boxed in red; inside to outside helices are boxed in blue (A). 11 conserved residues common to rhodopsin family GPCRs are denoted by asterisks (A). Interrogation of available nematode EST, genomic and transcriptomic datasets show that at least 12 of the 16 nematode species from clades IV and V which express flp-32 also express a homologue of the Gp-flp-32R (B), demonstrating inter-clade conservation of the flp-32 receptor. B. xylophilis: Bursaphelenchus xylophilus; G. pallida: Globodera pallida; G rostochiensis: Globodera rostochiensis; M. hapla: Meloidogyne hapla; M. incognita: Meloidogyne incognita; M. paranaensis: Meloidogyne paranaensis; R. similis: Radopholus similis; S. ratti: Strongyloides ratti; A. caninum: Ancylostoma caninum; A. cantonensis: Ancylostoma cantonensis; A. ceylanicum; Ancylostoma ceylanicum; C. elegans: Caenorhabditis elegans; H. polygyrus: Heligmosomoides polygyrus; N. americanus: Necator americanus; N. brasiliensis: Nippostrongylus brasiliensis; T. circumcincta: Teladorsagia circumcincta.
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Related In: Results  -  Collection

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getmorefigures.php?uid=PMC3585147&req=5

ppat-1003169-g007: A novel Gp-flp-32 receptor (Gp-flp-32R) is expressed in Globodera pallida J2s and is conserved across the nematode phylum.A novel Gp-flp-32R which shows close homology to the C. elegans VRFa receptor 1 (C26F1.6) is expressed in G. pallida (A). The 389 amino acid protein has seven transmembrane helices; outside to inside helices are boxed in red; inside to outside helices are boxed in blue (A). 11 conserved residues common to rhodopsin family GPCRs are denoted by asterisks (A). Interrogation of available nematode EST, genomic and transcriptomic datasets show that at least 12 of the 16 nematode species from clades IV and V which express flp-32 also express a homologue of the Gp-flp-32R (B), demonstrating inter-clade conservation of the flp-32 receptor. B. xylophilis: Bursaphelenchus xylophilus; G. pallida: Globodera pallida; G rostochiensis: Globodera rostochiensis; M. hapla: Meloidogyne hapla; M. incognita: Meloidogyne incognita; M. paranaensis: Meloidogyne paranaensis; R. similis: Radopholus similis; S. ratti: Strongyloides ratti; A. caninum: Ancylostoma caninum; A. cantonensis: Ancylostoma cantonensis; A. ceylanicum; Ancylostoma ceylanicum; C. elegans: Caenorhabditis elegans; H. polygyrus: Heligmosomoides polygyrus; N. americanus: Necator americanus; N. brasiliensis: Nippostrongylus brasiliensis; T. circumcincta: Teladorsagia circumcincta.
Mentions: Database mining facilitated the identification of a putative FLP-32 receptor in G. pallida, orthologous to the C. elegans VRFa receptor R1 (C26F1.6; see Fig. 7A). RACE PCR and sequencing confirmed the sequence of the putative G. pallida flp-32 receptor (Gp-flp-32R). Primers designed to confirm the open reading frame of Gp-flp-32R generated a 1,170 nucleotide cDNA sequence (GenBank accession number JQ685132), encoding a 389 aa protein (Fig. 7A). Gp-flp-32R encodes seven transmembrane helices and conserved residues at positions 48, 52, 76, 80, 136–138, 222, 273 and 318–320; the 136–138 sequence (DRF) is a common variation on the DRY motif at the cytosolic end of the third transmembrane helix and is common to rhodopsin-like GPCRs (see Fig. 7A). In a reciprocal tBLASTn search of the C. elegans non-redundant nucleotide and protein database, C. elegans C26F1.6 was returned as the top scoring hit (53% identity to Gp-flp-32R). Further interrogation of the G. pallida EST database (GenBank) and genome assembly (Wellcome Trust Sanger Institute November 2010 supercontig assembly) between August and October 2011 with C. elegans C26F1.6 did not reveal additional homologous transcripts. BLAST searches of all available nematode EST, genomic and transcriptomic resources identified 12 C26F1.6 GPCR homologues from the 16 species which express FLP-32 encoding transcripts, spanning clades IV and V (Fig. 7B).

Bottom Line: This study investigates the role of flp-32 in G. pallida and shows that: (i) Gp-flp-32 encodes the peptide AMRNALVRFamide; (ii) Gp-flp-32 is expressed in the brain and ventral nerve cord of G. pallida; (iii) migration rate increases in Gp-flp-32-silenced worms; (iv) the ability of G. pallida to infect potato plant root systems is enhanced in Gp-flp-32-silenced worms; (v) a novel putative Gp-flp-32 receptor (Gp-flp-32R) is expressed in G. pallida; and, (vi) Gp-flp-32R-silenced worms also display an increase in migration rate.This work demonstrates that Gp-flp-32 plays an intrinsic role in the modulation of locomotory behaviour in G. pallida and putatively interacts with at least one novel G-protein coupled receptor (Gp-flp-32R).This is the first functional characterisation of a parasitic nematode FLP-GPCR.

View Article: PubMed Central - PubMed

Affiliation: Molecular Biosciences-Parasitology, Institute for Global Food Security, School of Biological Sciences, Queen's University Belfast, Belfast, United Kingdom.

ABSTRACT
Restrictions on nematicide usage underscore the need for novel control strategies for plant pathogenic nematodes such as Globodera pallida (potato cyst nematode) that impose a significant economic burden on plant cultivation activities. The nematode neuropeptide signalling system is an attractive resource for novel control targets as it plays a critical role in sensory and motor functions. The FMRFamide-like peptides (FLPs) form the largest and most diverse family of neuropeptides in invertebrates, and are structurally conserved across nematode species, highlighting the utility of the FLPergic system as a broad-spectrum control target. flp-32 is expressed widely across nematode species. This study investigates the role of flp-32 in G. pallida and shows that: (i) Gp-flp-32 encodes the peptide AMRNALVRFamide; (ii) Gp-flp-32 is expressed in the brain and ventral nerve cord of G. pallida; (iii) migration rate increases in Gp-flp-32-silenced worms; (iv) the ability of G. pallida to infect potato plant root systems is enhanced in Gp-flp-32-silenced worms; (v) a novel putative Gp-flp-32 receptor (Gp-flp-32R) is expressed in G. pallida; and, (vi) Gp-flp-32R-silenced worms also display an increase in migration rate. This work demonstrates that Gp-flp-32 plays an intrinsic role in the modulation of locomotory behaviour in G. pallida and putatively interacts with at least one novel G-protein coupled receptor (Gp-flp-32R). This is the first functional characterisation of a parasitic nematode FLP-GPCR.

Show MeSH
Related in: MedlinePlus