Department of Bioscience and Biotechnology, Bio/Molecular Informatics Center, Konkuk University, Seoul, Korea.
Mol Cells. 2011 Feb;31(2):191-7. doi: 10.1007/s10059-011-0021-y. Epub 2010 Dec 22.
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.
IFE-1 是秀丽隐杆线虫中与 eIF4E 同源的五个蛋白之一,eIF4E 是翻译起始复合物 eIF4F 的 mRNA 5' 帽结合成分。IFE-1 的缺失会导致精子缺陷,这表明 IFE-1 调节了精子功能所需的一组特定基因。为了进一步了解 IFE-1 的分子功能,进行了蛋白质组分析,以寻找在 ife-1(ok1978)缺失突变体中相对于 fem-3(q20)对照表达下调的精子蛋白。选择 fem-3(q20)突变体背景是因为它只在限制温度下产生精子。对总虫体蛋白进行 2D-DIGE 分析,并用 MALDI-TOF 质谱进一步鉴定差异表达的蛋白斑点。在鉴定的蛋白中,发现 GSP-3 和主要精子蛋白(MSP)在 ife-1(ok1978)缺失突变体中显著下调。此外,gsp-3 的 RNAi 导致了类似 ife-1 的表型。这些结果表明,IFE-1 是一些精子特异性蛋白高效表达所必需的,ife-1 突变体的受精缺陷主要是由于 GSP-3 水平降低所致。