Phosphorylation or Mutation of the ERK2 Activation Loop Alters Oligonucleotide Binding

Andrea C. McReynolds, Aroon S. Karra, Yan Li, Elias Daniel Lopez, Adrian G. Turjanski, Elhadji Dioum, Kristina Lorenz, Elma Zaganjor, Steve Stippec, Kathleen McGlynn, Svetlana Earnest, Melanie H. Cobb

Research output: Contribution to journalArticle

6 Citations (Scopus)

Abstract

The mitogen-activated protein kinase ERK2 is able to elicit a wide range of context-specific responses to distinct stimuli, but the mechanisms underlying this versatility remain in question. Some cellular functions of ERK2 are mediated through regulation of gene expression. In addition to phosphorylating numerous transcriptional regulators, ERK2 is known to associate with chromatin and has been shown to bind oligonucleotides directly. ERK2 is activated by the upstream kinases MEK1/2, which phosphorylate both tyrosine 185 and threonine 183. ERK2 requires phosphorylation on both sites to be fully active. Some additional ERK2 phosphorylation sites have also been reported, including threonine 188. It has been suggested that this phospho form has distinct properties. We detected some ERK2 phosphorylated on T188 in bacterial preparations of ERK2 by mass spectrometry and further demonstrate that phosphomimetic substitution of this ERK2 residue impairs its kinase activity toward well-defined substrates and also affects its DNA binding. We used electrophoretic mobility shift assays with oligonucleotides derived from the insulin gene promoter and other regions to examine effects of phosphorylation and mutations on the binding of ERK2 to DNA. We show that ERK2 can bind oligonucleotides directly. Phosphorylation and mutations alter DNA binding and support the idea that signaling functions may be influenced through an alternate phosphorylation site.

Original languageEnglish (US)
Pages (from-to)1909-1917
Number of pages9
JournalBiochemistry
Volume55
Issue number12
DOIs
StatePublished - Mar 29 2016

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Phosphorylation
Oligonucleotides
Chemical activation
Mutation
Threonine
DNA
Phosphotransferases
Electrophoretic mobility
Gene Expression Regulation
Electrophoretic Mobility Shift Assay
Mitogen-Activated Protein Kinases
Genetic Promoter Regions
Gene expression
Chromatin
Mass spectrometry
Tyrosine
Assays
Mass Spectrometry
Substitution reactions
Genes

ASJC Scopus subject areas

  • Biochemistry

Cite this

McReynolds, A. C., Karra, A. S., Li, Y., Lopez, E. D., Turjanski, A. G., Dioum, E., ... Cobb, M. H. (2016). Phosphorylation or Mutation of the ERK2 Activation Loop Alters Oligonucleotide Binding. Biochemistry, 55(12), 1909-1917. https://doi.org/10.1021/acs.biochem.6b00096

Phosphorylation or Mutation of the ERK2 Activation Loop Alters Oligonucleotide Binding. / McReynolds, Andrea C.; Karra, Aroon S.; Li, Yan; Lopez, Elias Daniel; Turjanski, Adrian G.; Dioum, Elhadji; Lorenz, Kristina; Zaganjor, Elma; Stippec, Steve; McGlynn, Kathleen; Earnest, Svetlana; Cobb, Melanie H.

In: Biochemistry, Vol. 55, No. 12, 29.03.2016, p. 1909-1917.

Research output: Contribution to journalArticle

McReynolds, AC, Karra, AS, Li, Y, Lopez, ED, Turjanski, AG, Dioum, E, Lorenz, K, Zaganjor, E, Stippec, S, McGlynn, K, Earnest, S & Cobb, MH 2016, 'Phosphorylation or Mutation of the ERK2 Activation Loop Alters Oligonucleotide Binding', Biochemistry, vol. 55, no. 12, pp. 1909-1917. https://doi.org/10.1021/acs.biochem.6b00096
McReynolds AC, Karra AS, Li Y, Lopez ED, Turjanski AG, Dioum E et al. Phosphorylation or Mutation of the ERK2 Activation Loop Alters Oligonucleotide Binding. Biochemistry. 2016 Mar 29;55(12):1909-1917. https://doi.org/10.1021/acs.biochem.6b00096
McReynolds, Andrea C. ; Karra, Aroon S. ; Li, Yan ; Lopez, Elias Daniel ; Turjanski, Adrian G. ; Dioum, Elhadji ; Lorenz, Kristina ; Zaganjor, Elma ; Stippec, Steve ; McGlynn, Kathleen ; Earnest, Svetlana ; Cobb, Melanie H. / Phosphorylation or Mutation of the ERK2 Activation Loop Alters Oligonucleotide Binding. In: Biochemistry. 2016 ; Vol. 55, No. 12. pp. 1909-1917.
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