Proweller A, Butler J S
Department of Microbiology and Immunology, University of Rochester School of Medicine and Dentistry, New York 14642, USA.
J Biol Chem. 1996 May 3;271(18):10859-65. doi: 10.1074/jbc.271.18.10859.
Poly(A)-binding protein, the most abundant eukaryotic mRNP protein, is known primarily for its association with polyadenylate tails of mRNA. In the yeast, Saccharomyces cerevisiae, this protein (Pabp) was found to be essential for viability and has been implicated in models featuring roles in mRNA stability and as an enhancer of translation initiation. Although the mechanism of action is unknown, it is thought to require an activity to bind poly(A) tails and an additional capacity for an interaction with 60 S ribosomal subunits, perhaps via ribosomal protein L46 (Rpl46). We have found that a significant amount of Pabp in wild-type cells is not associated with polyribosome complexes. The remaining majority, which is found in these complexes, maintains its association even in yeast cells deficient in polyadenylated mRNA and/or Rpl46. These observations suggest that Pabp may not require interaction with poly(A) tails during translation. Further treatment of polyribosome lysates with agents known to differentially disrupt components of polyribosomes indicated that Pabp may require contact with some RNA component of the polyribosome, which could be either non-poly(A)-rich sequences of the translated mRNA or possibly a component of the ribosome. These findings suggest that Pabp may possess the ability to bind to ribosomes independently of its interaction with poly(A). We discuss these conclusions with respect to current models suggesting a multifunctional binding capacity of Pabp.
聚腺苷酸结合蛋白是真核生物中含量最丰富的mRNA结合蛋白,主要因其与mRNA的聚腺苷酸尾巴结合而为人所知。在酿酒酵母中,这种蛋白(Pabp)被发现对细胞存活至关重要,并且在mRNA稳定性及翻译起始增强子相关模型中发挥作用。尽管其作用机制尚不清楚,但一般认为它需要具备结合聚腺苷酸尾巴的活性以及与60S核糖体亚基相互作用的额外能力,可能是通过核糖体蛋白L46(Rpl46)来实现。我们发现野生型细胞中大量的Pabp并不与多核糖体复合物结合。而存在于这些复合物中的其余大部分Pabp,即使在缺乏聚腺苷酸化mRNA和/或Rpl46的酵母细胞中也能维持其结合状态。这些观察结果表明,Pabp在翻译过程中可能不需要与聚腺苷酸尾巴相互作用。用已知能不同程度破坏多核糖体组分的试剂对多核糖体裂解物进行进一步处理表明,Pabp可能需要与多核糖体的某些RNA组分接触,这些组分可能是翻译mRNA的非富含聚腺苷酸序列,也可能是核糖体的一个组分。这些发现表明,Pabp可能具有独立于其与聚腺苷酸相互作用而与核糖体结合的能力。我们结合当前认为Pabp具有多功能结合能力的模型来讨论这些结论。