Keidel Achim, Conti Elena, Falk Sebastian
Department of Structural Cell Biology, Max-Planck-Institute of Biochemistry, Martinsried, Germany.
Max Perutz Laboratories, Department of Structural and Computational Biology, University of Vienna, Vienna, Austria.
Methods Mol Biol. 2020;2062:491-513. doi: 10.1007/978-1-4939-9822-7_24.
The RNA exosome is a macromolecular machine that degrades a large variety of RNAs from their 3'-end. It comprises the major 3'-to-5' exonuclease in the cell, completely degrades erroneous and overly abundant RNAs, and is also involved in the precise processing of RNAs. To degrade transcripts both specifically and efficiently the exosome functions together with compartment-specific cofactors. In the yeast S. cerevisiae, the exosome associates with the Ski complex in the cytoplasm and with Mtr4 alone or with Mtr4 as part of the TRAMP complex in the nucleus. Here we describe how to produce, purify, and assemble the Ski and TRAMP complexes from S. cerevisiae.
RNA外切体是一种大分子机器，可从RNA的3'端降解多种RNA。它包含细胞中主要的3'至5'外切核酸酶，能完全降解错误的和过量的RNA，还参与RNA的精确加工。为了特异性且高效地降解转录本，外切体与特定区室的辅助因子协同发挥作用。在酿酒酵母中，外切体在细胞质中与Ski复合物结合，在细胞核中单独与Mtr4结合，或作为TRAMP复合物的一部分与Mtr4结合。在此，我们描述了如何从酿酒酵母中制备、纯化和组装Ski和TRAMP复合物。
