Identifying phosphoCTD-associating proteins.

Journal Article

The C-terminal repeat domain (CTD) of the largest subunit of RNA polymerase II is hyperphosphorylated during transcription elongation. The phosphoCTD is known to bind to a subset of RNA processing factors and to several other nuclear proteins, thereby positioning them to efficiently carry out their elongation-linked functions. The authors propose that additional phosphoCTD-associating proteins (PCAPs) exist and describe a systematic biochemical approach for identifying such proteins. A binding probe is generated by using yeast CTD kinase I to exhaustively phosphorylate a CTD fusion protein. This phosphoCTD is used to probe fractionated yeast or mammalian extracts in a Far Western protein interaction assay. Putative PCAPs are further purified and identified by mass spectrometry.

Full Text

Duke Authors

Cited Authors

  • Phatnani, HP; Greenleaf, AL

Published Date

  • 2004

Published In

Volume / Issue

  • 257 /

Start / End Page

  • 17 - 28

PubMed ID

  • 14769993

International Standard Serial Number (ISSN)

  • 1064-3745

Digital Object Identifier (DOI)

  • 10.1385/1-59259-750-5:017

Language

  • eng

Conference Location

  • United States