Purification of Messenger Ribonucleoprotein Particles via a Tagged Nascent Polypeptide.

ABSTRACT: The cytoplasmic fates of mRNAs are influenced by interactions between RNA-binding proteins and cis regulatory motifs. In the cytoplasm, mRNAs are present as messenger ribonucleoprotein particles, which include not only proteins that bind directly to the mRNA, but also additional proteins that are recruited via protein-protein interactions. Many labs have sought to purify such particles from cells, with limited success. We here describe a simple two-step procedure to purify actively translated mRNAs, with their associated proteins, from polysomes. We use a reporter mRNA that encodes a protein with three streptavidin binding peptides at the N-terminus. The polysomal reporter mRNA, with associated proteins, is purified via binding to a streptavidin matrix. The method takes four days, and can

SUBMITTER: Inchaustegui Gil DP 

PROVIDER: S-EPMC4726818 | biostudies-literature | 2016

REPOSITORIES: biostudies-literature