Polysome analysis for determining mRNA and ribosome association in Saccharomyces cerevisiae

Methods Enzymol. 2013:530:193-206. doi: 10.1016/B978-0-12-420037-1.00011-7.


The control of mRNA translation is vital for gene expression and it is regulated under various physiological and pathological conditions (Groppo and Richter, 2009). For example, under some physiological conditions, translational initiation is impaired. Therefore, the association of mRNA with ribosomes and/or ribosomal subunits can provide a powerful means to dissect specific aspects of posttranscriptional mRNA regulation. This protocol describes a technique used to determine ribosome occupancy on mRNA.

Keywords: Cell lysis; Grow yeast cells; Polysome analysis; Sedimentation analysis; Sucrose gradient.

MeSH terms

  • Cell Fractionation / methods
  • Centrifugation, Density Gradient / methods
  • Polyribosomes / metabolism*
  • RNA, Fungal / metabolism
  • RNA, Messenger / metabolism*
  • Ribosomes / metabolism*
  • Saccharomyces cerevisiae / cytology*
  • Saccharomyces cerevisiae / growth & development
  • Saccharomyces cerevisiae / metabolism
  • Sucrose / chemistry


  • RNA, Fungal
  • RNA, Messenger
  • Sucrose