Chung S, Eckrich M, Perrone-Bizzozero N, Kohn D T, Furneaux H
Program in Molecular Pharmacology and Therapeutics, Memorial Sloan Kettering Cancer Center, New York, New York 10021, USA.
J Biol Chem. 1997 Mar 7;272(10):6593-8. doi: 10.1074/jbc.272.10.6593.
Previous studies have identified three brain proteins (40, 65 and 95 kDa, respectively) that specifically bind to the 3'-untranslated region of GAP-43 mRNA. In this study, using a specific monoclonal antibody, we now show that the 40-kDa proteins are members of the Elav-like protein family. This family of specific RNA-binding proteins comprise three neural specific members called HuD, HuC, and Hel-N1. We have shown that purified recombinant HuD can bind with high affinity to GAP-43 mRNA. In addition, we have mapped the binding site to a highly conserved 26-nucleotide sequence within the regulatory element. The binding of HuD to this site is readily displaced by RNA oligonucleotides encoding other HuD binding sites. We also show that only the first and second RNA binding domains of HuD are required for selective binding to GAP-43 mRNA.
先前的研究已鉴定出三种脑蛋白（分别为40、65和95 kDa），它们能特异性结合GAP-43 mRNA的3'非翻译区。在本研究中，我们使用一种特异性单克隆抗体，现已证明40 kDa的蛋白是Elav样蛋白家族的成员。这个特异性RNA结合蛋白家族包含三个神经特异性成员，即HuD、HuC和Hel-N1。我们已证明纯化的重组HuD能以高亲和力结合GAP-43 mRNA。此外，我们已将结合位点定位到调控元件内一个高度保守的26核苷酸序列上。HuD与该位点的结合很容易被编码其他HuD结合位点的RNA寡核苷酸置换。我们还表明，HuD仅需其第一和第二个RNA结合结构域就能选择性结合GAP-43 mRNA。
