UV crosslinked mRNA-binding proteins captured from leaf mesophyll protoplasts.

ABSTRACT:

Background

The complexity of RNA regulation is one of the current frontiers in animal and plant molecular biology research. RNA-binding proteins (RBPs) are characteristically involved in post-transcriptional gene regulation through interaction with RNA. Recently, the mRNA-bound proteome of mammalian cell lines has been successfully cataloged using a new method called interactome capture. This method relies on UV crosslinking of proteins to RNA, purifying the mRNA using complementary oligo-dT beads and identifying the crosslinked proteins using mass spectrometry. We describe here an optimized system of mRNA interactome capture for Arabidopsis thaliana leaf mesophyll protoplasts, a cell type often used in functional cellular assays.

Results

We established the conditions

SUBMITTER: Zhang Z 

PROVIDER: S-EPMC5093948 | biostudies-literature | 2016

REPOSITORIES: biostudies-literature