Phosphorylation at serine 31 targets tyrosine hydroxylase to vesicles for transport along microtubules

Ana Jorge-Finnigan, Rune Kleppe, Kunwar Jung-KC, Ming Ying, Michael Marie, Ivan Rios-Mondragon, Michael Francis Salvatore, Jaakko Saraste, Aurora Martinez

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Abstract

Tyrosine hydroxylase (TH) catalyzes the conversion of L-tyrosine into L-DOPA, which is the rate-limiting step in the synthesis of catecholamines, such as dopamine, in dopaminergergic neurons. Low dopamine levels and death of the dopaminergic neurons are hallmarks of Parkinson's disease (PD), where α-synuclein is also a key player. TH is highly regulated, notably by phosphorylation of several Ser/Thr residues in the N-terminal tail. However, the functional role of TH phosphorylation at the Ser-31 site (THSer(P)-31) remains unclear. Here, we report that THSer(P)-31 co-distributes with the Golgi complex and synaptic-like vesicles in rat and human dopaminergic cells. We also found that the TH microsomal fraction content decreases after inhibition of cyclin-dependent kinase 5 (Cdk5) and ERK1/2. The cellular distribution of an overexpressed phospho-null mutant, TH1-S31A, was restricted to the soma of neuroblastoma cells, with decreased association with the microsomal fraction, whereas a phospho-mimic mutant, TH1-S31E, was distributed throughout the soma and neurites. TH1-S31E associated with vesicular monoamine transporter 2 (VMAT2) and α-synuclein in neuroblastoma cells, and endogenous THSer(P)-31 was detected in VMAT2- and α-synuclein- immunoprecipitated mouse brain samples. Microtubule disruption or co-transfection with α-synuclein A53T, a PD-associated mutation, caused TH1-S31E accumulation in the cell soma. Our results indicate that Ser-31 phosphorylation may regulate TH subcellular localization by enabling its transport along microtubules, notably toward the projection terminals. These findings disclose a new mechanism of TH regulation by phosphorylation and reveal its interaction with key players in PD, opening up new research avenues for better understanding dopamine synthesis in physiological and pathological states.

Original languageEnglish
Pages (from-to)14092-14107
Number of pages16
JournalJournal of Biological Chemistry
Volume292
Issue number34
DOIs
StatePublished - 25 Aug 2017

Fingerprint

Transport Vesicles
Phosphorylation
Tyrosine 3-Monooxygenase
Synucleins
Microtubules
Serine
Carisoprodol
Vesicular Monoamine Transport Proteins
Parkinson Disease
Dopamine
Neuroblastoma
Neurons
Cyclin-Dependent Kinase 5
Synaptic Vesicles
Dopaminergic Neurons
Golgi Apparatus
Neurites
Catecholamines
Transfection
Tyrosine

Cite this

Jorge-Finnigan, A., Kleppe, R., Jung-KC, K., Ying, M., Marie, M., Rios-Mondragon, I., ... Martinez, A. (2017). Phosphorylation at serine 31 targets tyrosine hydroxylase to vesicles for transport along microtubules. Journal of Biological Chemistry, 292(34), 14092-14107. https://doi.org/10.1074/jbc.M116.762344
Jorge-Finnigan, Ana ; Kleppe, Rune ; Jung-KC, Kunwar ; Ying, Ming ; Marie, Michael ; Rios-Mondragon, Ivan ; Salvatore, Michael Francis ; Saraste, Jaakko ; Martinez, Aurora. / Phosphorylation at serine 31 targets tyrosine hydroxylase to vesicles for transport along microtubules. In: Journal of Biological Chemistry. 2017 ; Vol. 292, No. 34. pp. 14092-14107.
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abstract = "Tyrosine hydroxylase (TH) catalyzes the conversion of L-tyrosine into L-DOPA, which is the rate-limiting step in the synthesis of catecholamines, such as dopamine, in dopaminergergic neurons. Low dopamine levels and death of the dopaminergic neurons are hallmarks of Parkinson's disease (PD), where α-synuclein is also a key player. TH is highly regulated, notably by phosphorylation of several Ser/Thr residues in the N-terminal tail. However, the functional role of TH phosphorylation at the Ser-31 site (THSer(P)-31) remains unclear. Here, we report that THSer(P)-31 co-distributes with the Golgi complex and synaptic-like vesicles in rat and human dopaminergic cells. We also found that the TH microsomal fraction content decreases after inhibition of cyclin-dependent kinase 5 (Cdk5) and ERK1/2. The cellular distribution of an overexpressed phospho-null mutant, TH1-S31A, was restricted to the soma of neuroblastoma cells, with decreased association with the microsomal fraction, whereas a phospho-mimic mutant, TH1-S31E, was distributed throughout the soma and neurites. TH1-S31E associated with vesicular monoamine transporter 2 (VMAT2) and α-synuclein in neuroblastoma cells, and endogenous THSer(P)-31 was detected in VMAT2- and α-synuclein- immunoprecipitated mouse brain samples. Microtubule disruption or co-transfection with α-synuclein A53T, a PD-associated mutation, caused TH1-S31E accumulation in the cell soma. Our results indicate that Ser-31 phosphorylation may regulate TH subcellular localization by enabling its transport along microtubules, notably toward the projection terminals. These findings disclose a new mechanism of TH regulation by phosphorylation and reveal its interaction with key players in PD, opening up new research avenues for better understanding dopamine synthesis in physiological and pathological states.",
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Jorge-Finnigan, A, Kleppe, R, Jung-KC, K, Ying, M, Marie, M, Rios-Mondragon, I, Salvatore, MF, Saraste, J & Martinez, A 2017, 'Phosphorylation at serine 31 targets tyrosine hydroxylase to vesicles for transport along microtubules', Journal of Biological Chemistry, vol. 292, no. 34, pp. 14092-14107. https://doi.org/10.1074/jbc.M116.762344

Phosphorylation at serine 31 targets tyrosine hydroxylase to vesicles for transport along microtubules. / Jorge-Finnigan, Ana; Kleppe, Rune; Jung-KC, Kunwar; Ying, Ming; Marie, Michael; Rios-Mondragon, Ivan; Salvatore, Michael Francis; Saraste, Jaakko; Martinez, Aurora.

In: Journal of Biological Chemistry, Vol. 292, No. 34, 25.08.2017, p. 14092-14107.

Research output: Contribution to journalArticleResearchpeer-review

TY - JOUR

T1 - Phosphorylation at serine 31 targets tyrosine hydroxylase to vesicles for transport along microtubules

AU - Jorge-Finnigan, Ana

AU - Kleppe, Rune

AU - Jung-KC, Kunwar

AU - Ying, Ming

AU - Marie, Michael

AU - Rios-Mondragon, Ivan

AU - Salvatore, Michael Francis

AU - Saraste, Jaakko

AU - Martinez, Aurora

PY - 2017/8/25

Y1 - 2017/8/25

N2 - Tyrosine hydroxylase (TH) catalyzes the conversion of L-tyrosine into L-DOPA, which is the rate-limiting step in the synthesis of catecholamines, such as dopamine, in dopaminergergic neurons. Low dopamine levels and death of the dopaminergic neurons are hallmarks of Parkinson's disease (PD), where α-synuclein is also a key player. TH is highly regulated, notably by phosphorylation of several Ser/Thr residues in the N-terminal tail. However, the functional role of TH phosphorylation at the Ser-31 site (THSer(P)-31) remains unclear. Here, we report that THSer(P)-31 co-distributes with the Golgi complex and synaptic-like vesicles in rat and human dopaminergic cells. We also found that the TH microsomal fraction content decreases after inhibition of cyclin-dependent kinase 5 (Cdk5) and ERK1/2. The cellular distribution of an overexpressed phospho-null mutant, TH1-S31A, was restricted to the soma of neuroblastoma cells, with decreased association with the microsomal fraction, whereas a phospho-mimic mutant, TH1-S31E, was distributed throughout the soma and neurites. TH1-S31E associated with vesicular monoamine transporter 2 (VMAT2) and α-synuclein in neuroblastoma cells, and endogenous THSer(P)-31 was detected in VMAT2- and α-synuclein- immunoprecipitated mouse brain samples. Microtubule disruption or co-transfection with α-synuclein A53T, a PD-associated mutation, caused TH1-S31E accumulation in the cell soma. Our results indicate that Ser-31 phosphorylation may regulate TH subcellular localization by enabling its transport along microtubules, notably toward the projection terminals. These findings disclose a new mechanism of TH regulation by phosphorylation and reveal its interaction with key players in PD, opening up new research avenues for better understanding dopamine synthesis in physiological and pathological states.

AB - Tyrosine hydroxylase (TH) catalyzes the conversion of L-tyrosine into L-DOPA, which is the rate-limiting step in the synthesis of catecholamines, such as dopamine, in dopaminergergic neurons. Low dopamine levels and death of the dopaminergic neurons are hallmarks of Parkinson's disease (PD), where α-synuclein is also a key player. TH is highly regulated, notably by phosphorylation of several Ser/Thr residues in the N-terminal tail. However, the functional role of TH phosphorylation at the Ser-31 site (THSer(P)-31) remains unclear. Here, we report that THSer(P)-31 co-distributes with the Golgi complex and synaptic-like vesicles in rat and human dopaminergic cells. We also found that the TH microsomal fraction content decreases after inhibition of cyclin-dependent kinase 5 (Cdk5) and ERK1/2. The cellular distribution of an overexpressed phospho-null mutant, TH1-S31A, was restricted to the soma of neuroblastoma cells, with decreased association with the microsomal fraction, whereas a phospho-mimic mutant, TH1-S31E, was distributed throughout the soma and neurites. TH1-S31E associated with vesicular monoamine transporter 2 (VMAT2) and α-synuclein in neuroblastoma cells, and endogenous THSer(P)-31 was detected in VMAT2- and α-synuclein- immunoprecipitated mouse brain samples. Microtubule disruption or co-transfection with α-synuclein A53T, a PD-associated mutation, caused TH1-S31E accumulation in the cell soma. Our results indicate that Ser-31 phosphorylation may regulate TH subcellular localization by enabling its transport along microtubules, notably toward the projection terminals. These findings disclose a new mechanism of TH regulation by phosphorylation and reveal its interaction with key players in PD, opening up new research avenues for better understanding dopamine synthesis in physiological and pathological states.

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Jorge-Finnigan A, Kleppe R, Jung-KC K, Ying M, Marie M, Rios-Mondragon I et al. Phosphorylation at serine 31 targets tyrosine hydroxylase to vesicles for transport along microtubules. Journal of Biological Chemistry. 2017 Aug 25;292(34):14092-14107. https://doi.org/10.1074/jbc.M116.762344