Function of RRM domains of Drosophila melanogaster ELAV: Rnp1 mutations and rrm domain replacements with ELAV family proteins and SXL.

Function of RRM domains of Drosophila melanogaster ELAV: Rnp1 mutations and rrm domain replacements with ELAV family proteins and SXL.
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果蝇 ELAV 的 RRM 结构域的功能:Rnp1 突变以及 ELAV 家族蛋白和 SXL 的 rrm 结构域替换。

DOI:
10.1093/genetics/155.4.1789
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发表时间:
2000
期刊:
影响因子:
3.3
通讯作者:
White,K
White,K
中科院分区:
生物学2区
文献类型:
--
作者:
Lisbin,MJ;Gordon,M;Yannoni,YM;White,K

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ELAV蛋白家族的成员含有三个高度保守的RNA识别基序(RRM)。ELAV是果蝇ELAV家族的成员之一,具有重要的生物学功能,主要分布在细胞核内。为了研究ELAV体内功能是否需要每个ELAV RRM的RNA结合特性,单独突变每个RRM的RNA结合中关键的氨基酸残基。严格的遗传互补试验表明,当突变蛋白是ELAV的唯一来源时,每个RRM的RNA结合能力是ELAV功能所必需的。为了评估每个结构域对ELAV功能的特异性程度以及哪些结构域可能执行相关ELAV蛋白的共同功能,我们用来自RBP 9的相应RRM替换ELAV RRM,D.与ELAV最同源的黑素胃蛋白; HuD,一种人ELAV家族蛋白;和SXL,尽管在进化上相关,但不是ELAV家族成员。该分析显示,RRM 3替代品功能齐全,但RRM 1和RRM 2替代品基本上无功能。在不太严格的条件下,来自SXL的RRM 1和RRM 2置换和来自RBP 9的RRM 1置换能够在突变型亚纯型ELAV蛋白存在下提供补充功能。
Members of the ELAV family of proteins contain three RNA recognition motifs (RRMs), which are highly conserved. ELAV, aDrosophila melanogastermember of this family, provides a vital function and exhibits a predominantly nuclear localization. To investigate if the RNA-binding property of each of the ELAV RRMs is required for ELAV'sin vivofunction, amino acid residues critical in RNA binding for each RRM were individually mutated. A stringent genetic complementation test revealed that when the mutant protein was the sole source of ELAV, RNA-binding ability of each RRM was essential to ELAV function. To assess the degree to which each domain was specific for ELAV function and which domains perhaps performed a function common to related ELAV proteins, we substituted an ELAV RRM with the corresponding RRM from RBP9, theD. melanogasterprotein most homologous to ELAV; HuD, a human ELAV family protein; and SXL, which, although evolutionarily related, is not an ELAV family member. This analysis revealed that RRM3 replacements were fully functional, but RRM1 and RRM2 replacements were largely nonfunctional. Under less stringent conditions RRM1 and RRM2 replacements from SXL and RRM1 replacement from RBP9 were able to provide supplemental function in the presence of a mutant hypomorphic ELAV protein.
果蝇 elav 基因座上的两个不同的温度敏感等位基因被同一色氨酸密码子的无义突变抑制。
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