Analysis of DrkA kinase's role in STATa activation

Yukika Saga, Yumi Iwade, Tsuyoshi Araki, Megumi Ishikawa, Takefumi Kawata (Lead / Corresponding author)

Research output: Contribution to journalArticle

Abstract

Dictyostelium STATa is a homologue of metazoan signal transducers and activators of transcription (STATs) and is important for morphogenesis. STATa is activated by phosphorylation on Tyr702 when cells are exposed to extracellular cAMP. Although two tyrosine kinase-like (TKL) proteins, Pyk2 and Pyk3, have been definitively identified as STATc kinases, no kinase is known for STATa activation. Based on homology to the previously identified tyrosine-selective TKLs, we identified DrkA, a member of the TKL family and the Dictyostelium receptor-like kinase (DRK) subfamily, as a candidate STATa kinase. The drkA gene is almost exclusively expressed in prestalk A (pstA) cells, where STATa is activated. Transient over-expression of DrkA increased STATa phosphorylation, although over-expression of the protein causes a severe growth defect and cell death. Furthermore, recombinant DrkA protein is auto-phosphorylated on tyrosine and threonine residues, and an in vitro kinase assay shows that DrkA can phosphorylate STATa on Tyr702 in a STATa-SH2 (phosphotyrosine binding) domain-dependent manner. These observations strongly suggest that DrkA is one of the key regulators of STATa tyrosine phosphorylation and is consistent with it being the kinase that directly activates STATa.

Original languageEnglish
Pages (from-to)422-435
Number of pages14
JournalGenes to Cells
Volume24
Issue number6
Early online date19 Apr 2019
DOIs
Publication statusPublished - 17 Jun 2019

Fingerprint

Phosphotransferases
Tyrosine
Dictyostelium
Phosphorylation
Protein-Tyrosine Kinases
Phosphotyrosine
Threonine
Transducers
Morphogenesis
Recombinant Proteins
Cell Death
Growth
Genes
Proteins

Keywords

  • Dictyostelium
  • cyclic AMP
  • phosphotyrosine signal
  • signal transducer and activator of transcription
  • tyrosine kinase-like kinase (TKL)

Cite this

Saga, Y., Iwade, Y., Araki, T., Ishikawa, M., & Kawata, T. (2019). Analysis of DrkA kinase's role in STATa activation. Genes to Cells, 24(6), 422-435. https://doi.org/10.1111/gtc.12686
Saga, Yukika ; Iwade, Yumi ; Araki, Tsuyoshi ; Ishikawa, Megumi ; Kawata, Takefumi. / Analysis of DrkA kinase's role in STATa activation. In: Genes to Cells. 2019 ; Vol. 24, No. 6. pp. 422-435.
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Saga, Y, Iwade, Y, Araki, T, Ishikawa, M & Kawata, T 2019, 'Analysis of DrkA kinase's role in STATa activation', Genes to Cells, vol. 24, no. 6, pp. 422-435. https://doi.org/10.1111/gtc.12686

Analysis of DrkA kinase's role in STATa activation. / Saga, Yukika; Iwade, Yumi; Araki, Tsuyoshi; Ishikawa, Megumi; Kawata, Takefumi (Lead / Corresponding author).

In: Genes to Cells, Vol. 24, No. 6, 17.06.2019, p. 422-435.

Research output: Contribution to journalArticle

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AU - Saga, Yukika

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AU - Araki, Tsuyoshi

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N2 - Dictyostelium STATa is a homologue of metazoan signal transducers and activators of transcription (STATs) and is important for morphogenesis. STATa is activated by phosphorylation on Tyr702 when cells are exposed to extracellular cAMP. Although two tyrosine kinase-like (TKL) proteins, Pyk2 and Pyk3, have been definitively identified as STATc kinases, no kinase is known for STATa activation. Based on homology to the previously identified tyrosine-selective TKLs, we identified DrkA, a member of the TKL family and the Dictyostelium receptor-like kinase (DRK) subfamily, as a candidate STATa kinase. The drkA gene is almost exclusively expressed in prestalk A (pstA) cells, where STATa is activated. Transient over-expression of DrkA increased STATa phosphorylation, although over-expression of the protein causes a severe growth defect and cell death. Furthermore, recombinant DrkA protein is auto-phosphorylated on tyrosine and threonine residues, and an in vitro kinase assay shows that DrkA can phosphorylate STATa on Tyr702 in a STATa-SH2 (phosphotyrosine binding) domain-dependent manner. These observations strongly suggest that DrkA is one of the key regulators of STATa tyrosine phosphorylation and is consistent with it being the kinase that directly activates STATa.

AB - Dictyostelium STATa is a homologue of metazoan signal transducers and activators of transcription (STATs) and is important for morphogenesis. STATa is activated by phosphorylation on Tyr702 when cells are exposed to extracellular cAMP. Although two tyrosine kinase-like (TKL) proteins, Pyk2 and Pyk3, have been definitively identified as STATc kinases, no kinase is known for STATa activation. Based on homology to the previously identified tyrosine-selective TKLs, we identified DrkA, a member of the TKL family and the Dictyostelium receptor-like kinase (DRK) subfamily, as a candidate STATa kinase. The drkA gene is almost exclusively expressed in prestalk A (pstA) cells, where STATa is activated. Transient over-expression of DrkA increased STATa phosphorylation, although over-expression of the protein causes a severe growth defect and cell death. Furthermore, recombinant DrkA protein is auto-phosphorylated on tyrosine and threonine residues, and an in vitro kinase assay shows that DrkA can phosphorylate STATa on Tyr702 in a STATa-SH2 (phosphotyrosine binding) domain-dependent manner. These observations strongly suggest that DrkA is one of the key regulators of STATa tyrosine phosphorylation and is consistent with it being the kinase that directly activates STATa.

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Saga Y, Iwade Y, Araki T, Ishikawa M, Kawata T. Analysis of DrkA kinase's role in STATa activation. Genes to Cells. 2019 Jun 17;24(6):422-435. https://doi.org/10.1111/gtc.12686