Activation of Pyk2/RAFTK induces tyrosine phosphorylation of α‐synuclein via Src‐family kinases

Nakamura, Takeshi; Yamashita, Hiroshi; Nagano, Yoshito; Avraham, Shalom; Avraham, Hava; Matsumoto, Masayasu; Nakamura, Shotaro

doi:10.1016/s0014-5793(02)02861-2

Cited by 37 publications

(36 citation statements)

References 20 publications

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“…In vivo Src contributes to the tyrosine phosphorylation of ASAP1 either by directly phosphorylating ASAP1 or by enhancing Pyk2 activity toward ASAP1, e.g. by an activating phosphorylation of Pyk2 as has been suggested previously (1,30,31).…”

Section: Asap1 Tyrosine Phosphorylation By Pyk2 Modulates Its Gap Actmentioning

confidence: 61%

The Tyrosine Kinase Pyk2 Regulates Arf1 Activity by Phosphorylation and Inhibition of the Arf-GTPase-activating Protein ASAP1

Kruljac‐Letunic

Moelleken

Kallin

et al. 2003

Journal of Biological Chemistry

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Proline-rich tyrosine kinase 2 (Pyk2), a non-receptor tyrosine kinase structurally related to focal adhesion kinase, has been implicated in the regulation of mitogen-activated protein kinase cascades and ion channels, the induction of apoptosis, and in the modulation of the cytoskeleton. In order to understand how Pyk2 signaling mediates these diverse cellular functions, we performed a yeast two-hybrid screening using the C-terminal part of Pyk2 that contains potential protein-protein interaction sites as bait. A prominent binder of Pyk2 identified by this method was the Arf-GTPase-activating protein ASAP1. Pyk2-ASAP1 interaction was confirmed in pull-down as well as in co-immunoprecipitation experiments, and contact sites were mapped to the proline-rich regions of Pyk2 and the SH3 domain of ASAP1. Pyk2 directly phosphorylates ASAP1 on tyrosine residues in vitro and increases ASAP1 tyrosine phosphorylation when co-expressed in HEK293T cells. Phosphorylation of tyrosine 308 and 782 affects the phosphoinositide binding profile of ASAP1, and fluorimetric Arf-GTPase assays with purified proteins revealed an inhibition of ASAP1 GTPase-activating protein activity by Pyk2-mediated tyrosine phosphorylation. We therefore provide evidence for a functional interaction between Pyk2 and ASAP1 and a regulation of ASAP1 and hence Arf1 activity by Pyk2-mediated tyrosine phosphorylation. Pyk2 and FAK1 comprise a distinct family of non-receptor protein tyrosine kinases that are involved in transmission of extracellular signals to intracellular kinase cascades and regulation of diverse cellular responses such as adhesion, proliferation, differentiation, and apoptosis (1). Pyk2 and FAK share about 45% amino acid identity and a similar domain structure: a unique N terminus containing a FERM (band 4

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Section: Asap1 Tyrosine Phosphorylation By Pyk2 Modulates Its Gap Actmentioning

confidence: 61%

The Tyrosine Kinase Pyk2 Regulates Arf1 Activity by Phosphorylation and Inhibition of the Arf-GTPase-activating Protein ASAP1

Kruljac‐Letunic

Moelleken

Kallin

et al. 2003

Journal of Biological Chemistry

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“…(µm hour -1 ) for each construct between 3 hours and 4.5 hours after glycerol shock (n=52-121 neurons). α-Synuclein mutants affect axonal transport Phosphorylation of α-synuclein is known to occur on serine residues 87 and 129 as well as tyrosine residues Pronin et al, 2000;Nakamura et al, 2001;Ellis et al, 2001;Negro et al, 2002;Nakamura et al, 2002). Here, we have demonstrated that mimicking protein phosphorylation by mutating specific serine residues to aspartate might have a direct influence on α-synuclein axonal transport as each of the S87D and S129D mutations reduced the rate of α-synuclein transport.…”

Section: Discussionmentioning

confidence: 78%

“…Phosphorylation of α-synuclein on serine residues diminishes its affinity for vesicles Pronin et al, 2000), suggesting that, as is likely to be the case for neurofilaments and microtubule-associated protein 1B, the phosphorylation state of α-synuclein might regulate its axonal transport (Jung et al, 2000). In addition to phosphorylation of α-synuclein on serine residues, tyrosine phosphorylation of α-synuclein has also been reported, both in vitro and in cells (Nakamura et al, 2001;Ellis et al, 2001;Negro et al, 2002;Nakamura et al, 2002). However, any effects of tyrosine or serine phosphorylation of α-synuclein on its axonal transport α-Synuclein is a major protein constituent of Lewy bodies and mutations in α-synuclein cause familial autosomal dominant Parkinson's disease.…”

mentioning

confidence: 98%

Parkinson's disease α-synuclein mutations exhibit defective axonal transport in cultured neurons

Saha

Hill

Utton

et al. 2004

Journal of Cell Science

161

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α-Synuclein is a major protein constituent of Lewy bodies and mutations in α-synuclein cause familial autosomal dominant Parkinson's disease. One explanation for the formation of perikaryal and neuritic aggregates of α-synuclein, which is a presynaptic protein, is that the mutations disrupt α-synuclein transport and lead to its proximal accumulation. We found that mutant forms of α-synuclein, either associated with Parkinson's disease (A30P or A53T) or mimicking defined serine, but not tyrosine, phosphorylation states exhibit reduced axonal transport following transfection into cultured neurons. Furthermore, transfection of A30P, but not wild-type, α-synuclein results in accumulation of the protein proximal to the cell body. We propose that the reduced axonal transport exhibited by the Parkinson's disease-associated α-synuclein mutants examined in this study might contribute to perikaryal accumulation of α-synuclein and hence Lewy body formation and neuritic abnormalities in diseased brain.

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“…Casein kinase 1 and 2 can also phosphorylate Ser-129 of ␣-synuclein in cultured cells (13). In addition, the ␣-synuclein Tyr-125 residue is phosphorylated by c-Src and Fyn (34,35). Although ␣-synuclein is constitutively phosphorylated at Ser-87 as well as at Ser-129 residues (13), the kinase targeting serine 87 residue and its physiological role have not been described.…”

Section: Discussionmentioning

confidence: 99%

Dyrk1A Phosphorylates α-Synuclein and Enhances Intracellular Inclusion Formation

Kim

Sung

Lee

et al. 2006

Journal of Biological Chemistry

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Lewy bodies (LBs) are pathological hallmarks of Parkinson disease (PD) but also occur in Alzheimer disease (AD) and dementia of LBs. ␣-Synuclein, the major component of LBs, is observed in the brain of Down syndrome (DS) patients with AD. Dyrk1A, a dual specificity tyrosine-regulated kinase (Dyrk) family member, is the mammalian ortholog of the Drosophila minibrain (Mnb) gene, essential for normal postembryonic neurogenesis. The Dyrk1A gene resides in the human chromosome 21q22.2 region, which is associated with DS anomalies, including mental retardation. In this study, we examined whether Dyrk1A interacts with ␣-synuclein and subsequently affects intracellular ␣-synuclein inclusion formation in immortalized hippocampal neuronal (H19-7) cells. Dyrk1A selectively binds to ␣-synuclein in transformed and primary neuronal cells. ␣-Synuclein overexpression, followed by basic fibroblast growth factor-induced neuronal differentiation, resulted in cell death. We observed that accompanying cell death was increased ␣-synuclein phosphorylation and intracytoplasmic aggregation. In addition, the transfection of kinase-inactive Dyrk1A or Dyrk1A small interfering RNA blocked ␣-synuclein phosphorylation and aggregate formation. In vitro kinase assay of anti-Dyrk1A immunocomplexes demonstrated that Dyrk1A could phosphorylate ␣-synuclein at Ser-87. Furthermore, aggregates formed by phosphorylated ␣-synuclein have a distinct morphology and are more neurotoxic compared with aggregates composed of unmodified wild type ␣-synuclein. These findings suggest ␣-synuclein inclusion formation regulated by Dyrk1A, potentially affecting neuronal cell viability.

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Activation of Pyk2/RAFTK induces tyrosine phosphorylation of α‐synuclein via Src‐family kinases

Cited by 37 publications

References 20 publications

The Tyrosine Kinase Pyk2 Regulates Arf1 Activity by Phosphorylation and Inhibition of the Arf-GTPase-activating Protein ASAP1

The Tyrosine Kinase Pyk2 Regulates Arf1 Activity by Phosphorylation and Inhibition of the Arf-GTPase-activating Protein ASAP1

Parkinson's disease α-synuclein mutations exhibit defective axonal transport in cultured neurons

Dyrk1A Phosphorylates α-Synuclein and Enhances Intracellular Inclusion Formation

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