PDF(Pubmed)
Abstract：
BACKGROUND: Ribosome biogenesis is the process of assembling ribosome complexes that regulate cell proliferation and differentiation with potential regulatory effects on development. Many factors regulate ribosome biological processes. Nin one binding protein (Nob1) has received widespread attention as key genes regulating ribosome biogenesis-the 3\' end of the 20S rRNA is cleaved by Nob1 at cleavage site D to form 18S rRNA, generating translationally capable 40S subunit. As a ribosome biogenesis factor, Nob1 may regulate the development of organisms, but almost nothing is known about the function of Nob1 for any parasitic nematode. We explored the functional role of NOBP-1 (the homologous gene of Nob1) encoding gene from a parasitic nematode-Strongyloides stercoralis.
METHODS: The full-length cDNA, gDNA and promoter region of Ss-nobp-1 was identified using protein BLAST in WormBase ParaSite according to the Caenorhabditis elegans NOBP-1 sequence to analyze the gene structure. RNA sequencing (RNA-seq) data in wormbase were retrieved and analyzed to assess the transcript abundance of Ss-nobp-1 in seven developmental stages of S. stercoralis. The standard method for gonadal microinjection of constructs was carried out to determine the anatomic expression patterns of Ss-nobp-1. The interaction between Ss-NOBP-1 and partner of NOBP-1 (Ss-PNO-1) was assessed by yeast two-hybridization and bimolecular fluorescence complementarity (BiFC) experiments.
RESULTS: The NOBP-1 encoding gene Ss-nopb-1 from the zoonotic parasite S. stercoralis has been isolated and characterized. The genomic DNA representing Ss-nobp-1 includes a 1599-bp coding region and encodes a protein comprising 403 amino acids (aa), which contains conserved PIN domain and zinc ribbon domain. RNA-seq analysis revealed that Ss-nobp-1 transcripts are present throughout the seven developmental stages in S. stercoralis and have higher transcription levels in iL3, L3 and P Female. Ss-nobp-1 is expressed mainly in the intestine of transgenic S. stercoralis larvae, and there is a direct interaction between Ss-NOBP-1 and Ss-PNO-1.
CONCLUSIONS: Collectively, Ss-NOBP-1 has a potential role in embryo formation and the infective process, and findings from this study provide a sound foundation for investigating its function during the development of parasitic nematode.
METHODS: The full-length cDNA, gDNA and promoter region of Ss-nobp-1 was identified using protein BLAST in WormBase ParaSite according to the Caenorhabditis elegans NOBP-1 sequence to analyze the gene structure. RNA sequencing (RNA-seq) data in wormbase were retrieved and analyzed to assess the transcript abundance of Ss-nobp-1 in seven developmental stages of S. stercoralis. The standard method for gonadal microinjection of constructs was carried out to determine the anatomic expression patterns of Ss-nobp-1. The interaction between Ss-NOBP-1 and partner of NOBP-1 (Ss-PNO-1) was assessed by yeast two-hybridization and bimolecular fluorescence complementarity (BiFC) experiments.
RESULTS: The NOBP-1 encoding gene Ss-nopb-1 from the zoonotic parasite S. stercoralis has been isolated and characterized. The genomic DNA representing Ss-nobp-1 includes a 1599-bp coding region and encodes a protein comprising 403 amino acids (aa), which contains conserved PIN domain and zinc ribbon domain. RNA-seq analysis revealed that Ss-nobp-1 transcripts are present throughout the seven developmental stages in S. stercoralis and have higher transcription levels in iL3, L3 and P Female. Ss-nobp-1 is expressed mainly in the intestine of transgenic S. stercoralis larvae, and there is a direct interaction between Ss-NOBP-1 and Ss-PNO-1.
CONCLUSIONS: Collectively, Ss-NOBP-1 has a potential role in embryo formation and the infective process, and findings from this study provide a sound foundation for investigating its function during the development of parasitic nematode.
摘要：
背景：核糖体生物发生是组装调节细胞增殖和分化的核糖体复合物的过程，对发育具有潜在的调节作用。许多因素调节核糖体生物过程。Nin一结合蛋白(Nob1)作为调控核糖体生物发生的关键基因受到广泛关注——20SrRNA的3'端在切割位点D被Nob1切割形成18SrRNA,生成具有翻译能力的40S亚基。作为核糖体生物发生因子，Nob1可能调节生物体的发育，但对Nob1对任何寄生线虫的功能几乎一无所知。我们探索了来自寄生线虫-类线虫的NOBP-1（Nob1的同源基因）编码基因的功能作用。
方法：全长cDNA，根据秀丽隐杆线虫NOBP-1序列，使用WormBaseParaSite中的BLAST蛋白鉴定了Ss-nobbp-1的gDNA和启动子区，以分析基因结构。检索并分析了蠕虫中的RNA测序（RNA-seq）数据，以评估S.stercoralis的七个发育阶段中Ss-nobp-1的转录本丰度。进行构建体的性腺显微注射的标准方法以确定Ss-nobp-1的解剖表达模式。通过酵母双杂交和双分子荧光互补（BiFC）实验评估了Ss-NOBP-1与NOBP-1伴侣（Ss-PNO-1）之间的相互作用。
结果：已分离并鉴定了人畜共患寄生虫S.stercoralis的NOBP-1编码基因Ss-nopb-1。代表Ss-nobbp-1的基因组DNA包括1599bp的编码区，并编码包含403个氨基酸（aa）的蛋白质，其中包含保守的PIN域和锌带域。RNA-seq分析显示，Ss-nobbp-1转录本存在于子宫链球菌的七个发育阶段，并且在iL3，L3和P雌性中具有更高的转录水平。Ss-nobbp-1主要在转基因胸骨链球菌幼虫的肠道中表达，Ss-NOBP-1和Ss-PNO-1之间存在直接相互作用。
结论：总的来说，Ss-NOBP-1在胚胎形成和感染过程中具有潜在的作用，这项研究的发现为研究其在寄生线虫发育过程中的功能提供了坚实的基础。
方法：全长cDNA，根据秀丽隐杆线虫NOBP-1序列，使用WormBaseParaSite中的BLAST蛋白鉴定了Ss-nobbp-1的gDNA和启动子区，以分析基因结构。检索并分析了蠕虫中的RNA测序（RNA-seq）数据，以评估S.stercoralis的七个发育阶段中Ss-nobp-1的转录本丰度。进行构建体的性腺显微注射的标准方法以确定Ss-nobp-1的解剖表达模式。通过酵母双杂交和双分子荧光互补（BiFC）实验评估了Ss-NOBP-1与NOBP-1伴侣（Ss-PNO-1）之间的相互作用。
结果：已分离并鉴定了人畜共患寄生虫S.stercoralis的NOBP-1编码基因Ss-nopb-1。代表Ss-nobbp-1的基因组DNA包括1599bp的编码区，并编码包含403个氨基酸（aa）的蛋白质，其中包含保守的PIN域和锌带域。RNA-seq分析显示，Ss-nobbp-1转录本存在于子宫链球菌的七个发育阶段，并且在iL3，L3和P雌性中具有更高的转录水平。Ss-nobbp-1主要在转基因胸骨链球菌幼虫的肠道中表达，Ss-NOBP-1和Ss-PNO-1之间存在直接相互作用。
结论：总的来说，Ss-NOBP-1在胚胎形成和感染过程中具有潜在的作用，这项研究的发现为研究其在寄生线虫发育过程中的功能提供了坚实的基础。
