Determining in vivo activity of the yeast cytoplasmic exosome

Methods Enzymol. 2008:448:227-39. doi: 10.1016/S0076-6879(08)02612-8.

Abstract

A 3'-exoribonuclease complex, termed the exosome, has important functions in the cytoplasm, as well as in the nucleus, and is involved in 3'-processing and/or decay of many RNAs. This chapter will discuss methods to study cytoplasmic exosome function in yeast with in vivo approaches. The first section will describe mutants that are available to study the processing or decay of a specific RNA by the nuclear or cytoplasmic exosome. The second section will discuss methods to determine whether the cytoplasmic exosome is functional under a specific condition(s) with reporter mRNAs that are known substrates of this complex.

MeSH terms

  • Exoribonucleases / analysis*
  • Exoribonucleases / metabolism*
  • Exosomes / genetics
  • Exosomes / metabolism*
  • Humans
  • Mutation / genetics
  • RNA / genetics
  • RNA / metabolism
  • Saccharomyces cerevisiae / cytology*
  • Saccharomyces cerevisiae / enzymology*
  • Transcription, Genetic / genetics

Substances

  • RNA
  • Exoribonucleases