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Regulation of sperm-specific proteins by IFE-1, a germline-specific homolog of eIF4E, in C. elegans

机译:秀丽隐杆线虫IFE-1(eIF4E的种系特异性同源物)对精子特异性蛋白的调节

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摘要

IFE-1 is one of the five C. elegans homologs of eIF4E, which is the mRNA 5′ cap-binding component of the translation initiation complex eIF4F. Depletion of IFE-1 causes defects in sperm, suggesting that IFE-1 regulates a subset of genes required for sperm functions. To further understand the molecular function of IFE-1, proteomic analysis was performed to search for sperm proteins that are downregulated in ife-1(ok1978); fem-3(q20) mutants relative to the fem-3(q20) control. The fem-3(q20) mutant background was used because it only produces sperm at restrictive temperature. Total worm proteins were subjected to 2D-DIGE, and differentially expressed protein spots were further identified by MALDI-TOF mass spectrometry. Among the identified proteins, GSP-3 and Major Sperm Proteins (MSPs) were found to be significantly down-regulated in the ife-1(ok1978) mutant. Moreover, RNAi of gsp-3 caused an ife-1-like phenotype. These results suggest that IFE-1 is required for efficient expression of some sperm-specific proteins, and the fertilization defect of ife-1 mutant is caused mainly by a reduced level of GSP-3.
机译:IFE-1是eIF4E的五个秀丽隐杆线虫同源物之一,它是翻译起始复合物eIF4F的mRNA 5'帽结合成分。 IFE-1的耗尽会导致精子缺陷,这表明IFE-1调节了精子功能所需基因的一个子集。为了进一步了解IFE-1的分子功能,进行了蛋白质组学分析以寻找在ife-1中被下调的精子蛋白(ok1978); fem-3(q20)突变体相对于fem-3(q20)对照。使用fem-3(q20)突变体背景是因为它仅在限制温度下产生精子。将总蠕虫蛋白质进行2D-DIGE处理,并通过MALDI-TOF质谱进一步鉴定差异表达的蛋白质斑点。在鉴定出的蛋白质中,发现ife-1(ok1978)突变体中GSP-3和主要精子蛋白质(MSP)明显下调。而且,gsp-3的RNAi引起了ife-1样表型。这些结果表明,IFE-1是某些精子特异性蛋白有效表达所必需的,而ife-1突变体的受精缺陷主要是由GSP-3水平降低引起的。

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