Genome-wide Mapping of 5'-monophosphorylated Ends of Mammalian Nascent RNA Transcripts.

ABSTRACT: In eukaryotic cells, RNA biogenesis generally requires processing of the nascent transcript as it is being synthesized by RNA polymerase. These processing events include endonucleolytic cleavage, exonucleolytic trimming, and splicing of the growing nascent transcript. Endonucleolytic cleavage events that generate an exposed 5'-monophosphorylated (5'-PO₄) end on the growing nascent transcript occur in the maturation of rRNAs, tRNAs, and mRNAs. These 5'-PO₄ ends can be a target of further processing or be subjected to 5'-3' exonucleolytic digestion that may result in termination of transcription. Here, we describe how to identify 5'-PO₄ ends of intermediates in nascent RNA metabolism. We capture these species via metabolic labeling with bromouridine followed

SUBMITTER: Cortazar MA 

PROVIDER: S-EPMC10518774 | biostudies-literature | 2023 Sep

REPOSITORIES: biostudies-literature