Substrate Specificity of the Kinase P-TEFb towards the RNA Polymerase II C-Terminal Domain

Eric B. Gibbs, Tatiana Nikolaevna Laremore, Grace A. Usher, Bede Portz, Erik C. Cook, Scott A. Showalter

Research output: Contribution to journalArticle

Abstract

The positive transcription elongation factor b (P-TEFb) promotes transcription elongation through phosphorylation of the RNA polymerase II C-terminal domain. This process is not well understood, partly due to difficulties in determining the specificity of P-TEFb toward the various heptad repeat motifs within the C-terminal domain. A simple assay using mass spectrometry was developed to identify the substrate specificity of the Drosophila melanogaster P-TEFb (DmP-TEFb) in vitro. This assay demonstrated that DmP-TEFb preferentially phosphorylates Ser5 and, surprisingly, that pre-phosphorylation or conserved amino acid variation at the 7-position in the heptad can alter DmP-TEFb specificity, leading to the creation of distinct double-phosphorylation marks.

Original languageEnglish (US)
Pages (from-to)1909-1911
Number of pages3
JournalBiophysical journal
Volume113
Issue number9
DOIs
StatePublished - Nov 7 2017

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RNA Polymerase III
Peptide Elongation Factors
RNA Polymerase II
Positive Transcriptional Elongation Factor B
Substrate Specificity
Drosophila melanogaster
Transcription Factors
Phosphotransferases
Phosphorylation
Mass Spectrometry
Amino Acids

All Science Journal Classification (ASJC) codes

  • Biophysics

Cite this

Gibbs, Eric B. ; Laremore, Tatiana Nikolaevna ; Usher, Grace A. ; Portz, Bede ; Cook, Erik C. ; Showalter, Scott A. / Substrate Specificity of the Kinase P-TEFb towards the RNA Polymerase II C-Terminal Domain. In: Biophysical journal. 2017 ; Vol. 113, No. 9. pp. 1909-1911.
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Substrate Specificity of the Kinase P-TEFb towards the RNA Polymerase II C-Terminal Domain. / Gibbs, Eric B.; Laremore, Tatiana Nikolaevna; Usher, Grace A.; Portz, Bede; Cook, Erik C.; Showalter, Scott A.

In: Biophysical journal, Vol. 113, No. 9, 07.11.2017, p. 1909-1911.

Research output: Contribution to journalArticle

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AB - The positive transcription elongation factor b (P-TEFb) promotes transcription elongation through phosphorylation of the RNA polymerase II C-terminal domain. This process is not well understood, partly due to difficulties in determining the specificity of P-TEFb toward the various heptad repeat motifs within the C-terminal domain. A simple assay using mass spectrometry was developed to identify the substrate specificity of the Drosophila melanogaster P-TEFb (DmP-TEFb) in vitro. This assay demonstrated that DmP-TEFb preferentially phosphorylates Ser5 and, surprisingly, that pre-phosphorylation or conserved amino acid variation at the 7-position in the heptad can alter DmP-TEFb specificity, leading to the creation of distinct double-phosphorylation marks.

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