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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

Transcriptional profiles of Ss-riok-1.Transcript abundances were determined for the C-terminal coding region of Ss-riok-1, in seven developmental stages: parasitic females (P Female), post-parasitic first-stage larvae (PP L1), post-parasitic third-stage larvae (PP L3), free-living females (FL Female), post free-living first-stage larvae (PFL L1), infectious third-stage larvae (iL3), and in vivo activated third-stage larvae (L3+). Transcript abundances were calculated as fragments per kilobase of coding exon per million mapped reads (FPKM). Bracket with 1 star represent the significant difference (p<0.05), brackets with 3 stars represent the significant difference (p<0.01) in transcript abundances between the two selected stages. Error bars represent 95% confidential intervals.
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pntd-0003062-g004: Transcriptional profiles of Ss-riok-1.Transcript abundances were determined for the C-terminal coding region of Ss-riok-1, in seven developmental stages: parasitic females (P Female), post-parasitic first-stage larvae (PP L1), post-parasitic third-stage larvae (PP L3), free-living females (FL Female), post free-living first-stage larvae (PFL L1), infectious third-stage larvae (iL3), and in vivo activated third-stage larvae (L3+). Transcript abundances were calculated as fragments per kilobase of coding exon per million mapped reads (FPKM). Bracket with 1 star represent the significant difference (p<0.05), brackets with 3 stars represent the significant difference (p<0.01) in transcript abundances between the two selected stages. Error bars represent 95% confidential intervals.

Mentions: Ss-riok-1-specific transcripts were detected in all developmental stages of S. stercoralis examined (Fig. 4). Abundance of these transcripts increases significantly during the transition from PFL L1 to iL3, and remains at a high level in the host-derived L3+. L3+ develop to the parthenogenetic P female during their migration in the host and reach the intestine; a significant decrease with the transcripts abundance of Ss-riok-1 (p<0.05) was found during migration and development. The reduced abundance of Ss-riok-1 transcripts during development of PP L1s to FL females was also detected. The abundance of Ss-riok-1 transcripts in iL3 is significantly greater than in PP L3 (p<0.001). By contrast, the abundance of Ss-riok-1 transcripts are significantly higher in P female and PP L1 than in FL female and PFL L1, respectively (p<0.001).


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)

Transcriptional profiles of Ss-riok-1.Transcript abundances were determined for the C-terminal coding region of Ss-riok-1, in seven developmental stages: parasitic females (P Female), post-parasitic first-stage larvae (PP L1), post-parasitic third-stage larvae (PP L3), free-living females (FL Female), post free-living first-stage larvae (PFL L1), infectious third-stage larvae (iL3), and in vivo activated third-stage larvae (L3+). Transcript abundances were calculated as fragments per kilobase of coding exon per million mapped reads (FPKM). Bracket with 1 star represent the significant difference (p<0.05), brackets with 3 stars represent the significant difference (p<0.01) in transcript abundances between the two selected stages. Error bars represent 95% confidential intervals.
© Copyright Policy
Related In: Results  -  Collection

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

pntd-0003062-g004: Transcriptional profiles of Ss-riok-1.Transcript abundances were determined for the C-terminal coding region of Ss-riok-1, in seven developmental stages: parasitic females (P Female), post-parasitic first-stage larvae (PP L1), post-parasitic third-stage larvae (PP L3), free-living females (FL Female), post free-living first-stage larvae (PFL L1), infectious third-stage larvae (iL3), and in vivo activated third-stage larvae (L3+). Transcript abundances were calculated as fragments per kilobase of coding exon per million mapped reads (FPKM). Bracket with 1 star represent the significant difference (p<0.05), brackets with 3 stars represent the significant difference (p<0.01) in transcript abundances between the two selected stages. Error bars represent 95% confidential intervals.
Mentions: Ss-riok-1-specific transcripts were detected in all developmental stages of S. stercoralis examined (Fig. 4). Abundance of these transcripts increases significantly during the transition from PFL L1 to iL3, and remains at a high level in the host-derived L3+. L3+ develop to the parthenogenetic P female during their migration in the host and reach the intestine; a significant decrease with the transcripts abundance of Ss-riok-1 (p<0.05) was found during migration and development. The reduced abundance of Ss-riok-1 transcripts during development of PP L1s to FL females was also detected. The abundance of Ss-riok-1 transcripts in iL3 is significantly greater than in PP L3 (p<0.001). By contrast, the abundance of Ss-riok-1 transcripts are significantly higher in P female and PP L1 than in FL female and PFL L1, respectively (p<0.001).

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