Duncan R, Etchison D, Hershey J W
J Biol Chem. 1983 Jun 10;258(11):7236-9.
Infection of HeLa cells by poliovirus results in the inhibition of translation of capped cellular mRNA. A plausible mechanism for this inhibition is that the structure of one or more initiation factors involved in the recognition of capped mRNA is altered. Eukaryotic initiation factor (eIF) 4A and eIF-4B are implicated in mRNA binding to 40 S ribosomal subunits and can be cross-linked to oxidized capped mRNA. We examined these factors in HeLa cell lysates by two-dimensional isoelectric focusing/sodium dodecyl sulfate-polyacrylamide gel electrophoresis and immunoblotting. No alterations in the number of molecules/cell, in the molecular size, or in extents of covalent modification were detected when lysates from infected and mock-infected cells were compared. The integrity of eIF-2 and several eIF-3 polypeptides was also examined and likewise no alterations were detected. The failure of the translational machinery to recognize capped mRNA therefore is not due to a change in the structure of these initiation factors.
脊髓灰质炎病毒感染HeLa细胞会导致对加帽细胞mRNA翻译的抑制。这种抑制的一种合理机制是参与识别加帽mRNA的一种或多种起始因子的结构发生了改变。真核起始因子(eIF)4A和eIF-4B与mRNA结合到40S核糖体亚基有关,并且可以与氧化的加帽mRNA交联。我们通过二维等电聚焦/十二烷基硫酸钠-聚丙烯酰胺凝胶电泳和免疫印迹法在HeLa细胞裂解物中检测了这些因子。当比较感染细胞和模拟感染细胞的裂解物时,未检测到每个细胞中分子数量、分子大小或共价修饰程度的改变。还检测了eIF-2和几种eIF-3多肽的完整性,同样未检测到改变。因此,翻译机制无法识别加帽mRNA并非由于这些起始因子的结构变化。