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NANOG is multiply phosphorylated and directly modified by ERK2 and CDK1 in vitro.

ABSTRACT: NANOG is a divergent homeobox protein and a core component of the transcriptional circuitry that sustains pluripotency and self-renewal. Although NANOG has been extensively studied on the transcriptional level, little is known regarding its posttranslational regulation, likely due to its low abundance and challenging physical properties. Here, we identify eleven phosphorylation sites on endogenous human NANOG, nine of which mapped to single amino acids. To screen for the signaling molecules that impart these modifications, we developed the multiplexed assay for kinase specificity (MAKS). MAKS simultaneously tests activity for up to ten kinases while directly identifying the substrate and exact site of phosphorylation. Using MAKS, we discovered site-specific phosphorylation by ERK2 and CDK1/CyclinA2, providing a putative link between key signaling pathways and NANOG.

SUBMITTER: Brumbaugh J

PROVIDER: S-EPMC3966117 | biostudies-literature | 2014 Jan

REPOSITORIES: biostudies-literature

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NANOG is multiply phosphorylated and directly modified by ERK2 and CDK1 in vitro.

Brumbaugh Justin J Russell Jason D JD Yu Pengzhi P Westphall Michael S MS Coon Joshua J JJ Thomson James A JA

Stem cell reports 20140114 1

NANOG is a divergent homeobox protein and a core component of the transcriptional circuitry that sustains pluripotency and self-renewal. Although NANOG has been extensively studied on the transcriptional level, little is known regarding its posttranslational regulation, likely due to its low abundance and challenging physical properties. Here, we identify eleven phosphorylation sites on endogenous human NANOG, nine of which mapped to single amino acids. To screen for the signaling molecules that ...[more]

PMID: 24678451

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