STAR Protocols (Dec 2020)

Differential Centrifugation to Enrich Bacterial Ribonucleoprotein Bodies (BR bodies) from Caulobacter crescentus

  • Nisansala S. Muthunayake,
  • Nadra Al-Husini,
  • Jared M. Schrader

DOI
https://doi.org/10.1016/j.xpro.2020.100205
Journal volume & issue
Vol. 1, no. 3
p. 100205

Abstract

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Summary: Bacterial RNP bodies (BR bodies) contain the mRNA decay machinery, but the collection of associated RNAs and proteins are poorly defined. Here, we present a protocol for the rapid differential centrifugation-based enrichment of BR bodies from Caulobacter crescentus cells. As native BR bodies are highly labile and dissociate by degrading internal mRNAs, an active site mutant of RNase E, which blocks dissolution of BR bodies, allows BR-body stabilization during enrichment.For complete details on the use and execution of this protocol, please refer to Al-Husini et al. (2020).

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