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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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Alignment of the inferred amino acid sequences of Strongyloides stercoralis Ss-RIOK-1 with RIOK-1s from nine other species.The nine selected species are Loa loa (XP_003135673.1, Ll-RIOK-1), Brugia malayi (EDP30009.1, Bm-RIOK-1), Caenorhabditis elegans (CCD67367.1, Ce-RIOK-1), Haemonchus contortus (ADW23592.1, Hc-RIOK-1), Trichostrogylus vitrinus (CAR64255.1, Tv-RIOK-1), Homo sapiens (NP_113668.2, Hs-RIOK-1), Mus musculus (NP_077204.2, Mm-RIOK-1), Danio rerio (NP_998160.1, Dr-RIOK-1), Arabidopsis thaliana (NP_180071.1, At-RIOK-1), Schistosoma mansoni (XP_002573653.1, Sm-RIOK-1). Alpha-helices A-I or beta-sheet structures are highlighted with light grey and marked under the alignment. The subdomains I-XI are marked above the alignment. Functional domains including ATP-binding motif (red), flexible loop (yellow), hinge (yellow), active site (red) and metal binding loop (yellow) are highlighted and labeled above the alignment. Asterisks indicate identical residues.
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pntd-0003062-g001: Alignment of the inferred amino acid sequences of Strongyloides stercoralis Ss-RIOK-1 with RIOK-1s from nine other species.The nine selected species are Loa loa (XP_003135673.1, Ll-RIOK-1), Brugia malayi (EDP30009.1, Bm-RIOK-1), Caenorhabditis elegans (CCD67367.1, Ce-RIOK-1), Haemonchus contortus (ADW23592.1, Hc-RIOK-1), Trichostrogylus vitrinus (CAR64255.1, Tv-RIOK-1), Homo sapiens (NP_113668.2, Hs-RIOK-1), Mus musculus (NP_077204.2, Mm-RIOK-1), Danio rerio (NP_998160.1, Dr-RIOK-1), Arabidopsis thaliana (NP_180071.1, At-RIOK-1), Schistosoma mansoni (XP_002573653.1, Sm-RIOK-1). Alpha-helices A-I or beta-sheet structures are highlighted with light grey and marked under the alignment. The subdomains I-XI are marked above the alignment. Functional domains including ATP-binding motif (red), flexible loop (yellow), hinge (yellow), active site (red) and metal binding loop (yellow) are highlighted and labeled above the alignment. Asterisks indicate identical residues.

Mentions: The full-length cDNA of Ss-riok-1 (GeneBank Accession No. KJ701282) is 1828 bp in length, including a 5′-UTR of 377 bp, a 17 bp 3′-UTR followed with the polyadenylation signal and a coding sequence of 1,434 bp encoding 477 amino acids. Neither a first nor a second spliced leader sequence (SL1 and SL2, respectively) was identified. In the protein sequence predicted from the gene, the RIOK-1 motif “LVHADLSEYNTL” [9] was identified (Fig. 1). The Ss-RIOK-1 shares high sequence identity (50–65%) to RIOK-1s from a diverse range of organisms, including vertebrates, amphibians, fish, plants and nematodes, with the highest identity (65%) to As-RIOK-1 from A. suum.


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)

Alignment of the inferred amino acid sequences of Strongyloides stercoralis Ss-RIOK-1 with RIOK-1s from nine other species.The nine selected species are Loa loa (XP_003135673.1, Ll-RIOK-1), Brugia malayi (EDP30009.1, Bm-RIOK-1), Caenorhabditis elegans (CCD67367.1, Ce-RIOK-1), Haemonchus contortus (ADW23592.1, Hc-RIOK-1), Trichostrogylus vitrinus (CAR64255.1, Tv-RIOK-1), Homo sapiens (NP_113668.2, Hs-RIOK-1), Mus musculus (NP_077204.2, Mm-RIOK-1), Danio rerio (NP_998160.1, Dr-RIOK-1), Arabidopsis thaliana (NP_180071.1, At-RIOK-1), Schistosoma mansoni (XP_002573653.1, Sm-RIOK-1). Alpha-helices A-I or beta-sheet structures are highlighted with light grey and marked under the alignment. The subdomains I-XI are marked above the alignment. Functional domains including ATP-binding motif (red), flexible loop (yellow), hinge (yellow), active site (red) and metal binding loop (yellow) are highlighted and labeled above the alignment. Asterisks indicate identical residues.
© Copyright Policy
Related In: Results  -  Collection

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

pntd-0003062-g001: Alignment of the inferred amino acid sequences of Strongyloides stercoralis Ss-RIOK-1 with RIOK-1s from nine other species.The nine selected species are Loa loa (XP_003135673.1, Ll-RIOK-1), Brugia malayi (EDP30009.1, Bm-RIOK-1), Caenorhabditis elegans (CCD67367.1, Ce-RIOK-1), Haemonchus contortus (ADW23592.1, Hc-RIOK-1), Trichostrogylus vitrinus (CAR64255.1, Tv-RIOK-1), Homo sapiens (NP_113668.2, Hs-RIOK-1), Mus musculus (NP_077204.2, Mm-RIOK-1), Danio rerio (NP_998160.1, Dr-RIOK-1), Arabidopsis thaliana (NP_180071.1, At-RIOK-1), Schistosoma mansoni (XP_002573653.1, Sm-RIOK-1). Alpha-helices A-I or beta-sheet structures are highlighted with light grey and marked under the alignment. The subdomains I-XI are marked above the alignment. Functional domains including ATP-binding motif (red), flexible loop (yellow), hinge (yellow), active site (red) and metal binding loop (yellow) are highlighted and labeled above the alignment. Asterisks indicate identical residues.
Mentions: The full-length cDNA of Ss-riok-1 (GeneBank Accession No. KJ701282) is 1828 bp in length, including a 5′-UTR of 377 bp, a 17 bp 3′-UTR followed with the polyadenylation signal and a coding sequence of 1,434 bp encoding 477 amino acids. Neither a first nor a second spliced leader sequence (SL1 and SL2, respectively) was identified. In the protein sequence predicted from the gene, the RIOK-1 motif “LVHADLSEYNTL” [9] was identified (Fig. 1). The Ss-RIOK-1 shares high sequence identity (50–65%) to RIOK-1s from a diverse range of organisms, including vertebrates, amphibians, fish, plants and nematodes, with the highest identity (65%) to As-RIOK-1 from A. suum.

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