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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 gene structure of Ss-riok-1 with comparison to its homologues from Caenorhabditis elegans and Haemonchus contortus.Black boxes indicate the exons, with the numbers above indicating the length of exon. Introns are indicated by slanted lines between the exons, with the numbers indicating the intron length. The 5′- and 3′- untranslated regions (UTR) of Ss-riok-1 and Ce-riok-1 are indicated with white boxes, with the numbers above the box indicating the length of the UTR.
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pntd-0003062-g003: The gene structure of Ss-riok-1 with comparison to its homologues from Caenorhabditis elegans and Haemonchus contortus.Black boxes indicate the exons, with the numbers above indicating the length of exon. Introns are indicated by slanted lines between the exons, with the numbers indicating the intron length. The 5′- and 3′- untranslated regions (UTR) of Ss-riok-1 and Ce-riok-1 are indicated with white boxes, with the numbers above the box indicating the length of the UTR.

Mentions: The genomic DNA representing Ss-riok-1 (GeneBank Accession No. KJ701282) is 5889 bp in length. The 377 bp 5′-UTR from cDNA is interrupted by two large introns of 711 bp and 3148 bp in length, respectively. The coding sequence of Ss-riok-1 encompasses 1,636 bp, containing three exons of 284–587 bp in size and two introns of 64 bp and 138 bp in size, respectively. The 17 bp 3′-UTR of Ss-riok-1 follows the third exon of the coding sequence (Fig. 3). Comparison with Ce-riok-1 (MO1B12.5a, sequences were retrieved from WormBase) from C. elegans and Hc-riok-1 from H. contortus[19] showed that these two homologues contain more introns than Ss-riok-1. The coding sequence of Ce-riok-1 contains eight exons of 72–532 bp in size and seven introns of 58–857 bp in size, whereas Hc-riok-1 has 16 exons of 61–200 bp in size and 15 introns of 30–520 bp in size [19].


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 gene structure of Ss-riok-1 with comparison to its homologues from Caenorhabditis elegans and Haemonchus contortus.Black boxes indicate the exons, with the numbers above indicating the length of exon. Introns are indicated by slanted lines between the exons, with the numbers indicating the intron length. The 5′- and 3′- untranslated regions (UTR) of Ss-riok-1 and Ce-riok-1 are indicated with white boxes, with the numbers above the box indicating the length of the UTR.
© Copyright Policy
Related In: Results  -  Collection

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

pntd-0003062-g003: The gene structure of Ss-riok-1 with comparison to its homologues from Caenorhabditis elegans and Haemonchus contortus.Black boxes indicate the exons, with the numbers above indicating the length of exon. Introns are indicated by slanted lines between the exons, with the numbers indicating the intron length. The 5′- and 3′- untranslated regions (UTR) of Ss-riok-1 and Ce-riok-1 are indicated with white boxes, with the numbers above the box indicating the length of the UTR.
Mentions: The genomic DNA representing Ss-riok-1 (GeneBank Accession No. KJ701282) is 5889 bp in length. The 377 bp 5′-UTR from cDNA is interrupted by two large introns of 711 bp and 3148 bp in length, respectively. The coding sequence of Ss-riok-1 encompasses 1,636 bp, containing three exons of 284–587 bp in size and two introns of 64 bp and 138 bp in size, respectively. The 17 bp 3′-UTR of Ss-riok-1 follows the third exon of the coding sequence (Fig. 3). Comparison with Ce-riok-1 (MO1B12.5a, sequences were retrieved from WormBase) from C. elegans and Hc-riok-1 from H. contortus[19] showed that these two homologues contain more introns than Ss-riok-1. The coding sequence of Ce-riok-1 contains eight exons of 72–532 bp in size and seven introns of 58–857 bp in size, whereas Hc-riok-1 has 16 exons of 61–200 bp in size and 15 introns of 30–520 bp in size [19].

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