Phosphorylation of cortactin by cyclin-dependent kinase 5 modulates actin bundling by the dynamin 1-cortactin ring-like complex and formation of filopodia and lamellipodia in NG108-15 glioma-derived cells

Tadashi Abe, The Mon La, Yuuzi Miyagaki, Eri Oya, Fan Yan Wei, Kento Sumida, Kenshiro Fujise, Tetsuya Takeda, Kazuhito Tomizawa, Kohji Takei, Hiroshi Yamada

Research output: Contribution to journalArticle

Abstract

Dynamin copolymerizes with cortactin to form a ring-like complex that bundles and stabilizes actin filaments. Actin bundle formation is crucial for generation of filopodia and lamellipodia, which guide migration, invasion, and metastasis of cancer cells. However, it is unknown how the dynamin-cortactin complex regulates actin bundle formation. The present study investigated phosphorylation of cortactin by cyclin-dependent kinase 5 (CDK5) and its effect on actin bundle formation by the dynamin-cortactin complex. CDK5 directly phosphorylated cortactin at T145/T219 in vitro. Phosphomimetic mutants in which one or both of these threonine residues was substituted by aspartate were used. The three phosphomimetic mutants (T145D, T219D and T145DT219D) had a decreased affinity for F-actin. Furthermore, electron microscopy demonstrated that these phosphomimetic mutants could not form a ring-like complex with dynamin 1. Consistently, the dynamin 1-phosphomimetic cortactin complexes exhibited decreased actin-bundling activity. Expression of the phosphomimetic mutants resulted in not only aberrant lamellipodia and short filopodia but also cell migration in NG108-15 glioma-derived cells. These results indicate that phosphorylation of cortactin by CDK5 regulates formation of lamellipodia and filopodia by modulating dynamin 1/cortactin-dependent actin bundling. Taken together, these findings suggest that CDK5 is a potential molecular target for anticancer therapy.

Original languageEnglish
Pages (from-to)550-558
Number of pages9
JournalInternational Journal of Oncology
Volume54
Issue number2
DOIs
Publication statusPublished - Feb 1 2019

Fingerprint

Dynamin I
Cortactin
Cyclin-Dependent Kinase 5
Pseudopodia
Glioma
Actins
Phosphorylation
Dynamins
Threonine
Actin Cytoskeleton
Aspartic Acid
Cell Movement
Electron Microscopy
Neoplasm Metastasis

Keywords

  • Actin
  • Bundle
  • Cortactin
  • Cyclin-dependent kinase 5
  • Dynamin

ASJC Scopus subject areas

  • Oncology
  • Cancer Research

Cite this

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title = "Phosphorylation of cortactin by cyclin-dependent kinase 5 modulates actin bundling by the dynamin 1-cortactin ring-like complex and formation of filopodia and lamellipodia in NG108-15 glioma-derived cells",
abstract = "Dynamin copolymerizes with cortactin to form a ring-like complex that bundles and stabilizes actin filaments. Actin bundle formation is crucial for generation of filopodia and lamellipodia, which guide migration, invasion, and metastasis of cancer cells. However, it is unknown how the dynamin-cortactin complex regulates actin bundle formation. The present study investigated phosphorylation of cortactin by cyclin-dependent kinase 5 (CDK5) and its effect on actin bundle formation by the dynamin-cortactin complex. CDK5 directly phosphorylated cortactin at T145/T219 in vitro. Phosphomimetic mutants in which one or both of these threonine residues was substituted by aspartate were used. The three phosphomimetic mutants (T145D, T219D and T145DT219D) had a decreased affinity for F-actin. Furthermore, electron microscopy demonstrated that these phosphomimetic mutants could not form a ring-like complex with dynamin 1. Consistently, the dynamin 1-phosphomimetic cortactin complexes exhibited decreased actin-bundling activity. Expression of the phosphomimetic mutants resulted in not only aberrant lamellipodia and short filopodia but also cell migration in NG108-15 glioma-derived cells. These results indicate that phosphorylation of cortactin by CDK5 regulates formation of lamellipodia and filopodia by modulating dynamin 1/cortactin-dependent actin bundling. Taken together, these findings suggest that CDK5 is a potential molecular target for anticancer therapy.",
keywords = "Actin, Bundle, Cortactin, Cyclin-dependent kinase 5, Dynamin",
author = "Tadashi Abe and La, {The Mon} and Yuuzi Miyagaki and Eri Oya and Wei, {Fan Yan} and Kento Sumida and Kenshiro Fujise and Tetsuya Takeda and Kazuhito Tomizawa and Kohji Takei and Hiroshi Yamada",
year = "2019",
month = "2",
day = "1",
doi = "10.3892/ijo.2018.4663",
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TY - JOUR

T1 - Phosphorylation of cortactin by cyclin-dependent kinase 5 modulates actin bundling by the dynamin 1-cortactin ring-like complex and formation of filopodia and lamellipodia in NG108-15 glioma-derived cells

AU - Abe, Tadashi

AU - La, The Mon

AU - Miyagaki, Yuuzi

AU - Oya, Eri

AU - Wei, Fan Yan

AU - Sumida, Kento

AU - Fujise, Kenshiro

AU - Takeda, Tetsuya

AU - Tomizawa, Kazuhito

AU - Takei, Kohji

AU - Yamada, Hiroshi

PY - 2019/2/1

Y1 - 2019/2/1

N2 - Dynamin copolymerizes with cortactin to form a ring-like complex that bundles and stabilizes actin filaments. Actin bundle formation is crucial for generation of filopodia and lamellipodia, which guide migration, invasion, and metastasis of cancer cells. However, it is unknown how the dynamin-cortactin complex regulates actin bundle formation. The present study investigated phosphorylation of cortactin by cyclin-dependent kinase 5 (CDK5) and its effect on actin bundle formation by the dynamin-cortactin complex. CDK5 directly phosphorylated cortactin at T145/T219 in vitro. Phosphomimetic mutants in which one or both of these threonine residues was substituted by aspartate were used. The three phosphomimetic mutants (T145D, T219D and T145DT219D) had a decreased affinity for F-actin. Furthermore, electron microscopy demonstrated that these phosphomimetic mutants could not form a ring-like complex with dynamin 1. Consistently, the dynamin 1-phosphomimetic cortactin complexes exhibited decreased actin-bundling activity. Expression of the phosphomimetic mutants resulted in not only aberrant lamellipodia and short filopodia but also cell migration in NG108-15 glioma-derived cells. These results indicate that phosphorylation of cortactin by CDK5 regulates formation of lamellipodia and filopodia by modulating dynamin 1/cortactin-dependent actin bundling. Taken together, these findings suggest that CDK5 is a potential molecular target for anticancer therapy.

AB - Dynamin copolymerizes with cortactin to form a ring-like complex that bundles and stabilizes actin filaments. Actin bundle formation is crucial for generation of filopodia and lamellipodia, which guide migration, invasion, and metastasis of cancer cells. However, it is unknown how the dynamin-cortactin complex regulates actin bundle formation. The present study investigated phosphorylation of cortactin by cyclin-dependent kinase 5 (CDK5) and its effect on actin bundle formation by the dynamin-cortactin complex. CDK5 directly phosphorylated cortactin at T145/T219 in vitro. Phosphomimetic mutants in which one or both of these threonine residues was substituted by aspartate were used. The three phosphomimetic mutants (T145D, T219D and T145DT219D) had a decreased affinity for F-actin. Furthermore, electron microscopy demonstrated that these phosphomimetic mutants could not form a ring-like complex with dynamin 1. Consistently, the dynamin 1-phosphomimetic cortactin complexes exhibited decreased actin-bundling activity. Expression of the phosphomimetic mutants resulted in not only aberrant lamellipodia and short filopodia but also cell migration in NG108-15 glioma-derived cells. These results indicate that phosphorylation of cortactin by CDK5 regulates formation of lamellipodia and filopodia by modulating dynamin 1/cortactin-dependent actin bundling. Taken together, these findings suggest that CDK5 is a potential molecular target for anticancer therapy.

KW - Actin

KW - Bundle

KW - Cortactin

KW - Cyclin-dependent kinase 5

KW - Dynamin

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U2 - 10.3892/ijo.2018.4663

DO - 10.3892/ijo.2018.4663

M3 - Article

VL - 54

SP - 550

EP - 558

JO - International Journal of Oncology

JF - International Journal of Oncology

SN - 1019-6439

IS - 2

ER -