Protein kinase A-dependent phosphorylation of Dock180 at serine residue 1250 is important for glioma growth and invasion stimulated by platelet derived-growth factor receptor α

Haizhong Feng, Yanxin Li, Yuhua Yin, Weiwei Zhang, Yanli Hou, Lei Zhang, Zuoqing Li, Baoshu Xie, Wei Qiang Gao, Jann N Sarkaria, Jeffery J. Raizer, C. David James, Andrew T. Parsa, Bo Hu, Shi Yuan Cheng

Research output: Contribution to journalArticle

7 Citations (Scopus)

Abstract

Background. Dedicator of cytokinesis 1 (Dock1 or Dock180), a bipartite guanine nucleotide exchange factor for Rac1, plays critical roles in receptor tyrosine kinase-stimulated cancer growth and invasion. Dock180 activity is required in cell migration cancer tumorigenesis promoted by platelet derived growth factor receptor (PDGFR) and epidermal growth factor receptor. Methods. To demonstrate whether PDGFRα promotes tumor malignant behavior through protein kinase A (PKA)-dependent serine phosphorylation of Dock180, we performed cell proliferation, viability, migration, immunoprecipitation, immunoblotting, colony formation, and in vivo tumorigenesis assays using established and short-term explant cultures of glioblastoma cell lines. Results. Stimulation of PDGFRα results in phosphorylation of Dock180 at serine residue 1250 (S1250), whereas PKA inhibitors H-89 and KT5720 oppose this phosphorylation. S1250 locates within the Rac1-binding Dock homology region 2 domain of Dock180, and its phosphorylation activates Rac1, p-Akt, and phosphorylated extracellular signal-regulated kinase 1/2, while promoting cell migration, in vitro. By expressing RNA interference (RNAi)-resistant wild-type Dock180, but not mutant Dock180 S1250L, we were able to rescue PDGFRα-associated signaling and biological activities in cultured glioblastoma multiforme (GBM) cells that had been treated with RNAi for suppression of endogenous Dock180. In addition, expression of the same RNAi-resistant Dock180 rescued an invasive phenotype of GBM cells following intracranial engraftment in immunocompromised mice. Conclusion. These data describe an important mechanism by which PDGFRα promotes glioma malignant phenotypes through PKA-dependent serine phosphorylation of Dock180, and the data thereby support targeting the PDGFRα-PKA-Dock180-Rac1 axis for treating GBM with molecular profiles indicating PDGFRα signaling dependency.

Original languageEnglish (US)
Pages (from-to)832-842
Number of pages11
JournalNeuro-Oncology
Volume17
Issue number6
DOIs
StatePublished - 2015

Fingerprint

Platelet-Derived Growth Factor Receptors
Cyclic AMP-Dependent Protein Kinases
Glioma
Serine
Phosphorylation
Glioblastoma
Growth
RNA Interference
Cell Movement
Carcinogenesis
Phenotype
Guanine Nucleotide Exchange Factors
Neoplasms
Cytokinesis
Mitogen-Activated Protein Kinase 3
Mitogen-Activated Protein Kinase 1
Receptor Protein-Tyrosine Kinases
Protein Kinase Inhibitors
Immunoprecipitation
Epidermal Growth Factor Receptor

Keywords

  • Dock180
  • Glioblastomas
  • PDGFRα
  • Phosphorylation
  • PKA

ASJC Scopus subject areas

  • Cancer Research
  • Oncology
  • Clinical Neurology

Cite this

Protein kinase A-dependent phosphorylation of Dock180 at serine residue 1250 is important for glioma growth and invasion stimulated by platelet derived-growth factor receptor α. / Feng, Haizhong; Li, Yanxin; Yin, Yuhua; Zhang, Weiwei; Hou, Yanli; Zhang, Lei; Li, Zuoqing; Xie, Baoshu; Gao, Wei Qiang; Sarkaria, Jann N; Raizer, Jeffery J.; James, C. David; Parsa, Andrew T.; Hu, Bo; Cheng, Shi Yuan.

In: Neuro-Oncology, Vol. 17, No. 6, 2015, p. 832-842.

Research output: Contribution to journalArticle

Feng, H, Li, Y, Yin, Y, Zhang, W, Hou, Y, Zhang, L, Li, Z, Xie, B, Gao, WQ, Sarkaria, JN, Raizer, JJ, James, CD, Parsa, AT, Hu, B & Cheng, SY 2015, 'Protein kinase A-dependent phosphorylation of Dock180 at serine residue 1250 is important for glioma growth and invasion stimulated by platelet derived-growth factor receptor α', Neuro-Oncology, vol. 17, no. 6, pp. 832-842. https://doi.org/10.1093/neuonc/nou323
Feng, Haizhong ; Li, Yanxin ; Yin, Yuhua ; Zhang, Weiwei ; Hou, Yanli ; Zhang, Lei ; Li, Zuoqing ; Xie, Baoshu ; Gao, Wei Qiang ; Sarkaria, Jann N ; Raizer, Jeffery J. ; James, C. David ; Parsa, Andrew T. ; Hu, Bo ; Cheng, Shi Yuan. / Protein kinase A-dependent phosphorylation of Dock180 at serine residue 1250 is important for glioma growth and invasion stimulated by platelet derived-growth factor receptor α. In: Neuro-Oncology. 2015 ; Vol. 17, No. 6. pp. 832-842.
@article{2913303edc6b44fb941a406a04ac0bf6,
title = "Protein kinase A-dependent phosphorylation of Dock180 at serine residue 1250 is important for glioma growth and invasion stimulated by platelet derived-growth factor receptor α",
abstract = "Background. Dedicator of cytokinesis 1 (Dock1 or Dock180), a bipartite guanine nucleotide exchange factor for Rac1, plays critical roles in receptor tyrosine kinase-stimulated cancer growth and invasion. Dock180 activity is required in cell migration cancer tumorigenesis promoted by platelet derived growth factor receptor (PDGFR) and epidermal growth factor receptor. Methods. To demonstrate whether PDGFRα promotes tumor malignant behavior through protein kinase A (PKA)-dependent serine phosphorylation of Dock180, we performed cell proliferation, viability, migration, immunoprecipitation, immunoblotting, colony formation, and in vivo tumorigenesis assays using established and short-term explant cultures of glioblastoma cell lines. Results. Stimulation of PDGFRα results in phosphorylation of Dock180 at serine residue 1250 (S1250), whereas PKA inhibitors H-89 and KT5720 oppose this phosphorylation. S1250 locates within the Rac1-binding Dock homology region 2 domain of Dock180, and its phosphorylation activates Rac1, p-Akt, and phosphorylated extracellular signal-regulated kinase 1/2, while promoting cell migration, in vitro. By expressing RNA interference (RNAi)-resistant wild-type Dock180, but not mutant Dock180 S1250L, we were able to rescue PDGFRα-associated signaling and biological activities in cultured glioblastoma multiforme (GBM) cells that had been treated with RNAi for suppression of endogenous Dock180. In addition, expression of the same RNAi-resistant Dock180 rescued an invasive phenotype of GBM cells following intracranial engraftment in immunocompromised mice. Conclusion. These data describe an important mechanism by which PDGFRα promotes glioma malignant phenotypes through PKA-dependent serine phosphorylation of Dock180, and the data thereby support targeting the PDGFRα-PKA-Dock180-Rac1 axis for treating GBM with molecular profiles indicating PDGFRα signaling dependency.",
keywords = "Dock180, Glioblastomas, PDGFRα, Phosphorylation, PKA",
author = "Haizhong Feng and Yanxin Li and Yuhua Yin and Weiwei Zhang and Yanli Hou and Lei Zhang and Zuoqing Li and Baoshu Xie and Gao, {Wei Qiang} and Sarkaria, {Jann N} and Raizer, {Jeffery J.} and James, {C. David} and Parsa, {Andrew T.} and Bo Hu and Cheng, {Shi Yuan}",
year = "2015",
doi = "10.1093/neuonc/nou323",
language = "English (US)",
volume = "17",
pages = "832--842",
journal = "Neuro-Oncology",
issn = "1522-8517",
publisher = "Oxford University Press",
number = "6",

}

TY - JOUR

T1 - Protein kinase A-dependent phosphorylation of Dock180 at serine residue 1250 is important for glioma growth and invasion stimulated by platelet derived-growth factor receptor α

AU - Feng, Haizhong

AU - Li, Yanxin

AU - Yin, Yuhua

AU - Zhang, Weiwei

AU - Hou, Yanli

AU - Zhang, Lei

AU - Li, Zuoqing

AU - Xie, Baoshu

AU - Gao, Wei Qiang

AU - Sarkaria, Jann N

AU - Raizer, Jeffery J.

AU - James, C. David

AU - Parsa, Andrew T.

AU - Hu, Bo

AU - Cheng, Shi Yuan

PY - 2015

Y1 - 2015

N2 - Background. Dedicator of cytokinesis 1 (Dock1 or Dock180), a bipartite guanine nucleotide exchange factor for Rac1, plays critical roles in receptor tyrosine kinase-stimulated cancer growth and invasion. Dock180 activity is required in cell migration cancer tumorigenesis promoted by platelet derived growth factor receptor (PDGFR) and epidermal growth factor receptor. Methods. To demonstrate whether PDGFRα promotes tumor malignant behavior through protein kinase A (PKA)-dependent serine phosphorylation of Dock180, we performed cell proliferation, viability, migration, immunoprecipitation, immunoblotting, colony formation, and in vivo tumorigenesis assays using established and short-term explant cultures of glioblastoma cell lines. Results. Stimulation of PDGFRα results in phosphorylation of Dock180 at serine residue 1250 (S1250), whereas PKA inhibitors H-89 and KT5720 oppose this phosphorylation. S1250 locates within the Rac1-binding Dock homology region 2 domain of Dock180, and its phosphorylation activates Rac1, p-Akt, and phosphorylated extracellular signal-regulated kinase 1/2, while promoting cell migration, in vitro. By expressing RNA interference (RNAi)-resistant wild-type Dock180, but not mutant Dock180 S1250L, we were able to rescue PDGFRα-associated signaling and biological activities in cultured glioblastoma multiforme (GBM) cells that had been treated with RNAi for suppression of endogenous Dock180. In addition, expression of the same RNAi-resistant Dock180 rescued an invasive phenotype of GBM cells following intracranial engraftment in immunocompromised mice. Conclusion. These data describe an important mechanism by which PDGFRα promotes glioma malignant phenotypes through PKA-dependent serine phosphorylation of Dock180, and the data thereby support targeting the PDGFRα-PKA-Dock180-Rac1 axis for treating GBM with molecular profiles indicating PDGFRα signaling dependency.

AB - Background. Dedicator of cytokinesis 1 (Dock1 or Dock180), a bipartite guanine nucleotide exchange factor for Rac1, plays critical roles in receptor tyrosine kinase-stimulated cancer growth and invasion. Dock180 activity is required in cell migration cancer tumorigenesis promoted by platelet derived growth factor receptor (PDGFR) and epidermal growth factor receptor. Methods. To demonstrate whether PDGFRα promotes tumor malignant behavior through protein kinase A (PKA)-dependent serine phosphorylation of Dock180, we performed cell proliferation, viability, migration, immunoprecipitation, immunoblotting, colony formation, and in vivo tumorigenesis assays using established and short-term explant cultures of glioblastoma cell lines. Results. Stimulation of PDGFRα results in phosphorylation of Dock180 at serine residue 1250 (S1250), whereas PKA inhibitors H-89 and KT5720 oppose this phosphorylation. S1250 locates within the Rac1-binding Dock homology region 2 domain of Dock180, and its phosphorylation activates Rac1, p-Akt, and phosphorylated extracellular signal-regulated kinase 1/2, while promoting cell migration, in vitro. By expressing RNA interference (RNAi)-resistant wild-type Dock180, but not mutant Dock180 S1250L, we were able to rescue PDGFRα-associated signaling and biological activities in cultured glioblastoma multiforme (GBM) cells that had been treated with RNAi for suppression of endogenous Dock180. In addition, expression of the same RNAi-resistant Dock180 rescued an invasive phenotype of GBM cells following intracranial engraftment in immunocompromised mice. Conclusion. These data describe an important mechanism by which PDGFRα promotes glioma malignant phenotypes through PKA-dependent serine phosphorylation of Dock180, and the data thereby support targeting the PDGFRα-PKA-Dock180-Rac1 axis for treating GBM with molecular profiles indicating PDGFRα signaling dependency.

KW - Dock180

KW - Glioblastomas

KW - PDGFRα

KW - Phosphorylation

KW - PKA

UR - http://www.scopus.com/inward/record.url?scp=84943181994&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=84943181994&partnerID=8YFLogxK

U2 - 10.1093/neuonc/nou323

DO - 10.1093/neuonc/nou323

M3 - Article

C2 - 25468898

AN - SCOPUS:84943181994

VL - 17

SP - 832

EP - 842

JO - Neuro-Oncology

JF - Neuro-Oncology

SN - 1522-8517

IS - 6

ER -