Macfarlane J, Müller M
Institut für Physikalische Biochemie, Universität München, Germany.
Eur J Biochem. 1995 Nov 1;233(3):766-71. doi: 10.1111/j.1432-1033.1995.766_3.x.
In eukaryotes, the cotranslational targeting of proteins to the endoplasmic reticular membrane is initially mediated by the signal-recognition particle (SRP), a ribonucleoprotein complex consisting of the 7SL RNA and six protein subunits. Since the discovery of sequence homology between (a) the Escherichia coli 4.5S RNA (Ffs) and 7SL RNA, and (b) the E. coli P48 (Ffh) and SRP 54-kDa subunit, more evidence has been obtained that E. coli also possesses an SRP-type pathway that acts in the translocation of secreted proteins. Such a pathway could possibly be involved in the cotranslational integration of hydrophobic membrane proteins that cannot be effectively targeted post-translationally due to folding and aggregation. In this study, we report that disruption of the E. coli SRP complex with a dominant lethal 4.5S RNA mutant in vivo prevents functional membrane integration of the E. coli lactose permease (LacY). Likewise, depletion of the P48 (Ffh) protein also results in a decrease in the amount of functional LacY inserted into the E. coli plasma membrane. In direct contrast, inhibition of SecA function does not affect LacY integration. These results suggest a major function of the bacterial SRP in the targeting and subsequent integration of hydrophobic membrane proteins as opposed to SecA mediating the post-translational targeting of secretory proteins.
在真核生物中,蛋白质共翻译靶向内质网膜最初由信号识别颗粒(SRP)介导,SRP是一种核糖核蛋白复合体,由7SL RNA和六个蛋白质亚基组成。自从发现(a)大肠杆菌4.5S RNA(Ffs)与7SL RNA之间以及(b)大肠杆菌P48(Ffh)与SRP 54-kDa亚基之间的序列同源性以来,更多证据表明大肠杆菌也拥有一种SRP型途径,该途径在分泌蛋白的转运中起作用。这样的途径可能参与了疏水膜蛋白的共翻译整合,这些疏水膜蛋白由于折叠和聚集而无法在翻译后有效地靶向。在本研究中,我们报告在体内用显性致死的4.5S RNA突变体破坏大肠杆菌SRP复合体可阻止大肠杆菌乳糖通透酶(LacY)的功能性膜整合。同样,P48(Ffh)蛋白的缺失也会导致插入大肠杆菌质膜的功能性LacY数量减少。与之形成直接对比的是,SecA功能的抑制并不影响LacY的整合。这些结果表明,细菌SRP在疏水膜蛋白的靶向和后续整合中具有主要功能,这与SecA介导分泌蛋白的翻译后靶向不同。