Sample Preparation for Mass Spectrometry-based Identification of RNA-binding Regions.

ABSTRACT: Noncoding RNAs play important roles in several nuclear processes, including regulating gene expression, chromatin structure, and DNA repair. In most cases, the action of noncoding RNAs is mediated by proteins whose functions are in turn regulated by these interactions with noncoding RNAs. Consistent with this, a growing number of proteins involved in nuclear functions have been reported to bind RNA and in a few cases the RNA-binding regions of these proteins have been mapped, often through laborious, candidate-based methods. Here, we report a detailed protocol to perform a high-throughput, proteome-wide unbiased identification of RNA-binding proteins and their RNA-binding regions. The methodology relies on the incorporation of a photoreactive uridine analog in the cellular RNA, followed by

SUBMITTER: Warneford-Thomson R 

PROVIDER: S-EPMC5752346 | biostudies-literature | 2017 Sep

REPOSITORIES: biostudies-literature