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旨在了解粪类圆线虫的一种RIO蛋白激酶编码基因Ss-RIOK-1的功能作用。

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

作者信息

Yuan Wang, Lok James B, Stoltzfus Jonathan D, Gasser Robin B, Fang Fang, Lei Wei-Qiang, Fang Rui, Zhou Yan-Qin, Zhao Jun-Long, Hu Min

机构信息

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.

Department of Pathobiology, School of Veterinary Medicine, University of Pennsylvania, Philadelphia, Pennsylvania, United States of America.

出版信息

PLoS Negl Trop Dis. 2014 Aug 7;8(8):e3062. doi: 10.1371/journal.pntd.0003062. eCollection 2014 Aug.

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.

摘要

背景

一些针对酿酒酵母和哺乳动物的研究表明,RIO蛋白激酶(RIOKs)参与核糖体生物合成、细胞周期进程和发育。然而,关于它们在寄生线虫中的功能,相关信息却很匮乏。我们旨在研究来自寄生于人和犬类的粪类圆线虫中RIOK-1编码基因的功能。

方法/主要发现:从粪类圆线虫中鉴定出RIOK-1蛋白编码基因Ss-riok-1。分离并测序了Ss-riok-1的全长cDNA、基因组DNA(gDNA)和推定的启动子区域。该cDNA包含1828 bp,包括一个377 bp的5'-非翻译区(UTR)、一个17 bp的3'-UTR和一个1434 bp的开放阅读框(ORF),编码一个含有477个氨基酸的蛋白质,该蛋白质含有一个RIOK-1特征基序。Ss-riok-1编码区的基因组序列长度为1636 bp,有三个外显子和两个内含子。推定的启动子区域包含4280 bp,含有保守的启动子元件,包括四个CAAT框、12个GATA框、八个E框(CANNTG)和38个TATA框。Ss-riok-1基因在所有发育阶段均有转录,在感染性第三期幼虫(iL3)中转录丰度最高。重组Ss-RIOK-1是一种活性激酶,能够进行磷酸化和自磷酸化。转基因粪类圆线虫幼虫中的转录报告基因表达模式表明,Ss-RIOK-1在头部、身体和尾部的神经元以及咽部和皮下组织中表达。

结论/意义:对编码基因的分子特征以及时空表达模式的研究,为RIOKs在寄生线虫生物学过程中的功能提供了初步线索。

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