Large-scale discovery of ERK2 substrates identifies ERK-mediated transcriptional regulation by ETV3.

TitleLarge-scale discovery of ERK2 substrates identifies ERK-mediated transcriptional regulation by ETV3.
Publication TypeJournal Article
Year of Publication2011
AuthorsCarlson, SM, Chouinard, CR, Labadorf, A, Lam, CJ, Schmelzle, K, Fraenkel, E, White, FM
JournalSci Signal
Volume4
Issue196
Paginationrs11
Date Published2011
ISSN1937-9145
Keywords3T3-L1 Cells, Adenosine Triphosphate, Amino Acid Motifs, Amino Acid Sequence, Animals, Binding Sites, Blotting, Western, Butadienes, Enzyme Inhibitors, Gene Expression Regulation, HEK293 Cells, Humans, Mass Spectrometry, Mice, Mitogen-Activated Protein Kinase 1, Mitogen-Activated Protein Kinases, Molecular Sequence Data, Mutation, Nitriles, Phosphorylation, Protein Binding, Protein Engineering, Repressor Proteins, Substrate Specificity
Abstract

The mitogen-activated protein kinase (MAPK) extracellular signal-regulated kinase 2 (ERK2) is ubiquitously expressed in mammalian tissues and is involved in a wide range of biological processes. Although MAPKs have been intensely studied, identification of their substrates remains challenging. We have optimized a chemical genetic system using analog-sensitive ERK2, a form of ERK2 engineered to use an analog of adenosine 5'-triphosphate (ATP), to tag and isolate ERK2 substrates in vitro. This approach identified 80 proteins phosphorylated by ERK2, 13 of which are known ERK2 substrates. The 80 substrates are associated with diverse cellular processes, including regulation of transcription and translation, mRNA processing, and regulation of the activity of the Rho family guanosine triphosphatases. We found that one of the newly identified substrates, ETV3 (a member of the E twenty-six family of transcriptional regulators), was extensively phosphorylated on sites within canonical and noncanonical ERK motifs. Phosphorylation of ETV3 regulated transcription by preventing its binding to DNA at promoters for several thousand genes, including some involved in negative feedback regulation of itself and of upstream signals.

DOI10.1126/scisignal.2002010
Alternate JournalSci Signal
PubMed ID22028470
PubMed Central IDPMC3779841
Grant ListES002109 / ES / NIEHS NIH HHS / United States
P30 CA014051 / CA / NCI NIH HHS / United States
P30 ES002109 / ES / NIEHS NIH HHS / United States
R01 CA118705 / CA / NCI NIH HHS / United States
R01 DK042816 / DK / NIDDK NIH HHS / United States
R01CA118705 / CA / NCI NIH HHS / United States
R01DK42816 / DK / NIDDK NIH HHS / United States
U54 CA112967 / CA / NCI NIH HHS / United States
U54CA112967 / CA / NCI NIH HHS / United States