Richter J D, Lorenz L J, Audet R G
J Biol Chem. 1985 Apr 10;260(7):4448-54.
Messenger RNA injected Xenopus oocytes exhibit a differential capacity for translation. mRNAs translated in the free cytoplasm are translated efficiently whereas mRNAs translated on the rough endoplasmic reticulum (RER) membrane are translated inefficiently. If mRNA injected oocytes are injected additionally with proteins isolated from the RER, enhanced translation of RER-bound mRNAs is observed. When examined by sucrose gradient centrifugation and RNA dot blots, most of the injected RER-bound mRNA sediments less than or equal to the 80 S monosome. The RER proteins recruit these preinitiated mRNAs onto polysomes as evidenced by a shift in sedimentation to the polysome region of a sucrose gradient. When examined by immunoblotting, the RER proteins are shown to contain a protein which reacts specifically with an antibody directed against docking protein (SRP-receptor protein). However, this putative docking protein does not appear to be the protein which actually recruits the preinitiated mRNAs onto polysomes.
注射到非洲爪蟾卵母细胞中的信使核糖核酸(mRNA)表现出不同的翻译能力。在游离细胞质中翻译的mRNA能高效翻译,而在糙面内质网(RER)膜上翻译的mRNA则翻译效率低下。如果向注射了mRNA的卵母细胞中额外注射从RER分离的蛋白质,就会观察到与RER结合的mRNA的翻译增强。通过蔗糖梯度离心和RNA斑点印迹法检测时,大多数注射的与RER结合的mRNA沉降速度小于或等于80S单体核糖体。RER蛋白将这些起始前的mRNA募集到多核糖体上,蔗糖梯度沉降至多核糖体区域的变化证明了这一点。通过免疫印迹法检测时,RER蛋白显示含有一种能与针对对接蛋白(信号识别颗粒受体蛋白)的抗体发生特异性反应的蛋白质。然而,这种假定的对接蛋白似乎并不是实际将起始前的mRNA募集到多核糖体上的蛋白质。