Simon R, Richter J D
Worcester Foundation For Experimental Biology, Shrewsbury, Massachusetts 01545.
Mol Cell Biol. 1990 Nov;10(11):5609-15. doi: 10.1128/mcb.10.11.5609-5615.1990.
The adenovirus E1A gene product is a potent transcriptional activator and nuclear oncoprotein. Like other regulatory proteins, E1A has a short half-life, in the range of 30 to 120 min. This short half-life, which was measured in cells synthesizing E1A, is not observed in cells injected with E1A protein made in bacteria or in vitro. In these cases, E1A is essentially refractory to degradation. In an attempt to reconcile this apparent paradox, we suggested that E1A was marked for degradation during its synthesis. Furthermore, we showed that a domain in the amino terminus of E1A was required for rapid degradation in cells translating E1A mRNA (J. M. Slavicek, N. C. Jones, and J. D. Richter, EMBO J. 7:3171-3180, 1988). In this study, we have used Xenopus laevis oocytes injected with mRNAs encoding altered E1A proteins to show that the amino-terminal tetrapeptide Met-Arg-His-Ile is required for E1A degradation. Even conservative amino acid substitutions in this degradation sequence render it nonfunctional. This degradation sequence can function as a transferable signal, since it induces instability when fused to another normally stable protein. Furthermore, the degradation sequence requires a proximity of no more than six residues from the amino terminus for activity. These data suggest that a trans-acting factor recognizes the amino terminus of E1A during the translation of its message to mark the protein for subsequent destruction.
腺病毒E1A基因产物是一种强效转录激活因子和核癌蛋白。与其他调节蛋白一样,E1A半衰期较短,在30至120分钟范围内。在合成E1A的细胞中测得的这种短半衰期,在注射细菌或体外制备的E1A蛋白的细胞中并未观察到。在这些情况下,E1A基本上不易降解。为了解决这一明显的矛盾,我们提出E1A在合成过程中被标记以便降解。此外,我们表明E1A氨基末端的一个结构域是细胞中翻译E1A mRNA时快速降解所必需的(J.M. Slavicek、N.C. Jones和J.D. Richter,《欧洲分子生物学组织杂志》7:3171 - 3180,1988年)。在本研究中,我们利用注射了编码改变的E1A蛋白的mRNA的非洲爪蟾卵母细胞,表明E1A降解需要氨基末端四肽Met - Arg - His - Ile。即使在这个降解序列中进行保守的氨基酸替换也会使其失去功能。这个降解序列可以作为一个可转移的信号,因为当它与另一种通常稳定的蛋白质融合时会诱导不稳定性。此外,降解序列从氨基末端起距离不超过六个残基时才能发挥作用。这些数据表明,一种反式作用因子在E1A信使RNA翻译过程中识别其氨基末端,从而标记该蛋白以便随后被破坏。