Mapping the Phosphorylation pattern of Drosophila melanogaster RNA polymerase II carboxyl-terminal domain using ultraviolet photodissociation mass spectrometry

Joshua E. Mayfield, Michelle R. Robinson, Victoria C. Cotham, Seema Irani, Wendy L. Matthews, Anjana Ram, David S. Gilmour, Joe R. Cannon, Yan Jessie Zhang, Jennifer S. Brodbelt

Research output: Contribution to journalArticle

12 Scopus citations

Abstract

Phosphorylation of the C-terminal domain of RNA polymerase II (CTD) plays an essential role in eukaryotic transcription by recruiting transcriptional regulatory factors to the active polymerase. However, the scarcity of basic residues and repetitive nature of the CTD sequence impose a huge challenge for site-specific characterization of phosphorylation, hindering our understanding of this crucial biological process. Herein, we apply LC-UVPD-MS methods to analyze posttranslational modification along native sequence CTDs. Application of our method to the Drosophila melanogaster CTD reveals the phosphorylation pattern of this model organism for the first time. The divergent nature of fly CTD allows us to derive rules defining how flanking residues affect phosphorylation choice by CTD kinases. Our data support the use of LC-UVPD-MS to decipher the CTD code and determine rules that program its function.

Original languageEnglish (US)
Pages (from-to)153-162
Number of pages10
JournalACS chemical biology
Volume12
Issue number1
DOIs
StatePublished - 2017

All Science Journal Classification (ASJC) codes

  • Biochemistry
  • Molecular Medicine

Fingerprint Dive into the research topics of 'Mapping the Phosphorylation pattern of Drosophila melanogaster RNA polymerase II carboxyl-terminal domain using ultraviolet photodissociation mass spectrometry'. Together they form a unique fingerprint.

  • Cite this

    Mayfield, J. E., Robinson, M. R., Cotham, V. C., Irani, S., Matthews, W. L., Ram, A., Gilmour, D. S., Cannon, J. R., Zhang, Y. J., & Brodbelt, J. S. (2017). Mapping the Phosphorylation pattern of Drosophila melanogaster RNA polymerase II carboxyl-terminal domain using ultraviolet photodissociation mass spectrometry. ACS chemical biology, 12(1), 153-162. https://doi.org/10.1021/acschembio.6b00729