Cell-type specific polysome profiling from mammalian tissues

Joseph Seimetz, Waqar Arif, Sushant Bangru, Mikel Hernaez, Auinash Kalsotra

Research output: Contribution to journalArticle

Abstract

The regulation of gene expression occurs through complex relationships between transcription, processing, turnover, and translation, which are only beginning to be elucidated. We know that at least for certain messenger (m) RNAs, processing, modifications, and sequence elements can greatly influence their translational output through recognition by translation and turn-over machinery. Recently, we and others have combined high-throughput sequencing technologies with traditional biochemical methods of studying translation to extend our understanding of these relationships. Additionally, there is growing importance given to how these processes may be regulated across varied cell types as a means to achieve tissue-specific expression of proteins. Here, we provide an in-depth methodology for polysome profiling to dissect the composition of mRNAs and proteins that make up the translatome from both whole tissues and a specific cell type isolated from mammalian tissue. Also, we provide a detailed computational workflow for the analysis of the next-generation sequencing data generated from these experiments.

Original languageEnglish (US)
Pages (from-to)131-139
Number of pages9
JournalMethods
Volume155
DOIs
StatePublished - Feb 15 2019

Keywords

  • Cell-type isolation from tissues
  • Next-generation sequencing and bioinformatics
  • Polysome profiling
  • Ribosomal occupancy shift
  • Translation regulation

ASJC Scopus subject areas

  • Molecular Biology
  • Biochemistry, Genetics and Molecular Biology(all)

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