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Toward understanding the functional role of Ss-RIOK-1, a RIO protein kinase-encoding gene of Strongyloides stercoralis.

Yuan W, Lok JB, Stoltzfus JD, Gasser RB, Fang F, Lei WQ, Fang R, Zhou YQ, Zhao JL, Hu M - PLoS Negl Trop Dis (2014)

Bottom Line: Recombinant Ss-RIOK-1 is an active kinase, capable of both phosphorylation and auto-phosphorylation.Patterns of transcriptional reporter expression in transgenic S. stercoralis larvae indicated that Ss-RIOK-1 is expressed in neurons of the head, body and tail as well as in pharynx and hypodermis.The characterization of the molecular and the temporal and spatial expression patterns of the encoding gene provide first clues as to functions of RIOKs in the biological processes of parasitic nematodes.

View Article: PubMed Central - PubMed

Affiliation: State Key Laboratory of Agricultural Microbiology, Key Laboratory of Development of Veterinary Diagnostic Products, Ministry of Agriculture, College of Veterinary Medicine, Huazhong Agricultural University, Wuhan, China.

ABSTRACT

Background: Some studies of Saccharomyces cerevisiae and mammals have shown that RIO protein kinases (RIOKs) are involved in ribosome biogenesis, cell cycle progression and development. However, there is a paucity of information on their functions in parasitic nematodes. We aimed to investigate the function of RIOK-1 encoding gene from Strongyloides stercoralis, a nematode parasitizing humans and dogs.

Methodology/principal findings: The RIOK-1 protein-encoding gene Ss-riok-1 was characterized from S. stercoralis. The full-length cDNA, gDNA and putative promoter region of Ss-riok-1 were isolated and sequenced. The cDNA comprises 1,828 bp, including a 377 bp 5'-UTR, a 17 bp 3'-UTR and a 1,434 bp ORF encoding a protein of 477 amino acids containing a RIOK-1 signature motif. The genomic sequence of the Ss-riok-1 coding region is 1,636 bp in length and has three exons and two introns. The putative promoter region comprises 4,280 bp and contains conserved promoter elements, including four CAAT boxes, 12 GATA boxes, eight E-boxes (CANNTG) and 38 TATA boxes. The Ss-riok-1 gene is transcribed throughout all developmental stages with the highest transcript abundance in the infective third-stage larva (iL3). Recombinant Ss-RIOK-1 is an active kinase, capable of both phosphorylation and auto-phosphorylation. Patterns of transcriptional reporter expression in transgenic S. stercoralis larvae indicated that Ss-RIOK-1 is expressed in neurons of the head, body and tail as well as in pharynx and hypodermis.

Conclusions/significance: The characterization of the molecular and the temporal and spatial expression patterns of the encoding gene provide first clues as to functions of RIOKs in the biological processes of parasitic nematodes.

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Related in: MedlinePlus

The Neighbor-joining tree of Strongyloides stercoralis Ss-RIOK-1 with 27 homologues from 23 selected species.These species contain nine nematode species, two plant species, two insects, three fish and amphibian species and six mammalian species. The RIOK-1 from Saccharomyces cerevisiae (CAA99317.1) is used as the outgroup. GenBank accession numbers of the homologous sequences are listed beside the species name. Bootstrap values are displayed above or below the branches.
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pntd-0003062-g002: The Neighbor-joining tree of Strongyloides stercoralis Ss-RIOK-1 with 27 homologues from 23 selected species.These species contain nine nematode species, two plant species, two insects, three fish and amphibian species and six mammalian species. The RIOK-1 from Saccharomyces cerevisiae (CAA99317.1) is used as the outgroup. GenBank accession numbers of the homologous sequences are listed beside the species name. Bootstrap values are displayed above or below the branches.

Mentions: Results of phylogenetic analyses (Fig. 2) showed that there is concordance in topology among the MP, ML and NJ trees. Ss-RIOK-1 groups with orthologues from clade V nematodes [47] with strong (99%) nodal support. The RIOK-1s from parasitic nematodes representing clade III [47] grouped together with strong (95%) support; all nematode RIOK-1s formed a cluster with absolute support to the exclusion of 17 RIOK-1s from 13 non-nematode species. Among the 17 RIOK-1s representing 13 non-nematode species, RIOK-1s from plants, mammals or insects each grouped together, respectively with high bootstrap support (99–100%). The RIOK-1s from other vertebrates, including fish and amphibians, grouped with the RIOK-1s from mammals with strong bootstrap support respectively (100%).


Toward understanding the functional role of Ss-RIOK-1, a RIO protein kinase-encoding gene of Strongyloides stercoralis.

Yuan W, Lok JB, Stoltzfus JD, Gasser RB, Fang F, Lei WQ, Fang R, Zhou YQ, Zhao JL, Hu M - PLoS Negl Trop Dis (2014)

The Neighbor-joining tree of Strongyloides stercoralis Ss-RIOK-1 with 27 homologues from 23 selected species.These species contain nine nematode species, two plant species, two insects, three fish and amphibian species and six mammalian species. The RIOK-1 from Saccharomyces cerevisiae (CAA99317.1) is used as the outgroup. GenBank accession numbers of the homologous sequences are listed beside the species name. Bootstrap values are displayed above or below the branches.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4125297&req=5

pntd-0003062-g002: The Neighbor-joining tree of Strongyloides stercoralis Ss-RIOK-1 with 27 homologues from 23 selected species.These species contain nine nematode species, two plant species, two insects, three fish and amphibian species and six mammalian species. The RIOK-1 from Saccharomyces cerevisiae (CAA99317.1) is used as the outgroup. GenBank accession numbers of the homologous sequences are listed beside the species name. Bootstrap values are displayed above or below the branches.
Mentions: Results of phylogenetic analyses (Fig. 2) showed that there is concordance in topology among the MP, ML and NJ trees. Ss-RIOK-1 groups with orthologues from clade V nematodes [47] with strong (99%) nodal support. The RIOK-1s from parasitic nematodes representing clade III [47] grouped together with strong (95%) support; all nematode RIOK-1s formed a cluster with absolute support to the exclusion of 17 RIOK-1s from 13 non-nematode species. Among the 17 RIOK-1s representing 13 non-nematode species, RIOK-1s from plants, mammals or insects each grouped together, respectively with high bootstrap support (99–100%). The RIOK-1s from other vertebrates, including fish and amphibians, grouped with the RIOK-1s from mammals with strong bootstrap support respectively (100%).

Bottom Line: Recombinant Ss-RIOK-1 is an active kinase, capable of both phosphorylation and auto-phosphorylation.Patterns of transcriptional reporter expression in transgenic S. stercoralis larvae indicated that Ss-RIOK-1 is expressed in neurons of the head, body and tail as well as in pharynx and hypodermis.The characterization of the molecular and the temporal and spatial expression patterns of the encoding gene provide first clues as to functions of RIOKs in the biological processes of parasitic nematodes.

View Article: PubMed Central - PubMed

Affiliation: State Key Laboratory of Agricultural Microbiology, Key Laboratory of Development of Veterinary Diagnostic Products, Ministry of Agriculture, College of Veterinary Medicine, Huazhong Agricultural University, Wuhan, China.

ABSTRACT

Background: Some studies of Saccharomyces cerevisiae and mammals have shown that RIO protein kinases (RIOKs) are involved in ribosome biogenesis, cell cycle progression and development. However, there is a paucity of information on their functions in parasitic nematodes. We aimed to investigate the function of RIOK-1 encoding gene from Strongyloides stercoralis, a nematode parasitizing humans and dogs.

Methodology/principal findings: The RIOK-1 protein-encoding gene Ss-riok-1 was characterized from S. stercoralis. The full-length cDNA, gDNA and putative promoter region of Ss-riok-1 were isolated and sequenced. The cDNA comprises 1,828 bp, including a 377 bp 5'-UTR, a 17 bp 3'-UTR and a 1,434 bp ORF encoding a protein of 477 amino acids containing a RIOK-1 signature motif. The genomic sequence of the Ss-riok-1 coding region is 1,636 bp in length and has three exons and two introns. The putative promoter region comprises 4,280 bp and contains conserved promoter elements, including four CAAT boxes, 12 GATA boxes, eight E-boxes (CANNTG) and 38 TATA boxes. The Ss-riok-1 gene is transcribed throughout all developmental stages with the highest transcript abundance in the infective third-stage larva (iL3). Recombinant Ss-RIOK-1 is an active kinase, capable of both phosphorylation and auto-phosphorylation. Patterns of transcriptional reporter expression in transgenic S. stercoralis larvae indicated that Ss-RIOK-1 is expressed in neurons of the head, body and tail as well as in pharynx and hypodermis.

Conclusions/significance: The characterization of the molecular and the temporal and spatial expression patterns of the encoding gene provide first clues as to functions of RIOKs in the biological processes of parasitic nematodes.

Show MeSH
Related in: MedlinePlus