The Arabidopsis mitogen-activated protein kinase 6 is associated with γ-tubulin on microtubules, phosphorylates EB1c and maintains spindle orientation under nitrosative stress

Lucie Kohoutová, Hana Kourová, Szilvia K. Nagy, Jindřich Volc, Petr Halada, T. Mészáros, Irute Meskiene, László Bögre, Pavla Binarová

Research output: Contribution to journalArticle

13 Citations (Scopus)

Abstract

Stress-activated plant mitogen-activated protein (MAP) kinase pathways play roles in growth adaptation to the environment by modulating cell division through cytoskeletal regulation, but the mechanisms are poorly understood. We performed protein interaction and phosphorylation experiments with cytoskeletal proteins, mass spectrometric identification of MPK6 complexes and immunofluorescence analyses of the microtubular cytoskeleton of mitotic cells using wild-type, mpk6-2 mutant and plants overexpressing the MAP kinase-inactivating phosphatase, AP2C3. We showed that MPK6 interacted with γ-tubulin and co-sedimented with plant microtubules polymerized in vitro. It was the active form of MAP kinase that was enriched with microtubules and followed similar dynamics to γ-tubulin, moving from poles to midzone during the anaphase-to-telophase transition. We found a novel substrate for MPK6, the microtubule plus end protein, EB1c. The mpk6-2 mutant was sensitive to 3-nitro-l-tyrosine (NO2-Tyr) treatment with respect to mitotic abnormalities, and root cells overexpressing AP2C3 showed defects in chromosome segregation and spindle orientation. Our data suggest that the active form of MAP kinase interacts with γ-tubulin on specific subsets of mitotic microtubules during late mitosis. MPK6 phosphorylates EB1c, but not EB1a, and has a role in maintaining regular planes of cell division under stress conditions.

Original languageEnglish
Pages (from-to)1061-1074
Number of pages14
JournalNew Phytologist
Volume207
Issue number4
DOIs
Publication statusPublished - Sep 1 2015

Fingerprint

Mitogen-Activated Protein Kinase 6
Tubulin
tubulin
mitogen-activated protein kinase
Arabidopsis
Microtubules
microtubules
Mitogen-Activated Protein Kinases
Cell Division
cell division
Mitogen-Activated Protein Kinase Phosphatases
Telophase
cytoskeletal proteins
telophase
mutants
Chromosome Segregation
Anaphase
Cytoskeletal Proteins
chromosome segregation
anaphase

Keywords

  • Arabidopsis
  • Cell division
  • EB1c
  • Microtubules
  • Mitogen-activated protein kinase
  • MPK6
  • Nitrosative stress
  • γ-tubulin

ASJC Scopus subject areas

  • Plant Science
  • Physiology

Cite this

The Arabidopsis mitogen-activated protein kinase 6 is associated with γ-tubulin on microtubules, phosphorylates EB1c and maintains spindle orientation under nitrosative stress. / Kohoutová, Lucie; Kourová, Hana; Nagy, Szilvia K.; Volc, Jindřich; Halada, Petr; Mészáros, T.; Meskiene, Irute; Bögre, László; Binarová, Pavla.

In: New Phytologist, Vol. 207, No. 4, 01.09.2015, p. 1061-1074.

Research output: Contribution to journalArticle

Kohoutová, Lucie ; Kourová, Hana ; Nagy, Szilvia K. ; Volc, Jindřich ; Halada, Petr ; Mészáros, T. ; Meskiene, Irute ; Bögre, László ; Binarová, Pavla. / The Arabidopsis mitogen-activated protein kinase 6 is associated with γ-tubulin on microtubules, phosphorylates EB1c and maintains spindle orientation under nitrosative stress. In: New Phytologist. 2015 ; Vol. 207, No. 4. pp. 1061-1074.
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AU - Nagy, Szilvia K.

AU - Volc, Jindřich

AU - Halada, Petr

AU - Mészáros, T.

AU - Meskiene, Irute

AU - Bögre, László

AU - Binarová, Pavla

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AB - Stress-activated plant mitogen-activated protein (MAP) kinase pathways play roles in growth adaptation to the environment by modulating cell division through cytoskeletal regulation, but the mechanisms are poorly understood. We performed protein interaction and phosphorylation experiments with cytoskeletal proteins, mass spectrometric identification of MPK6 complexes and immunofluorescence analyses of the microtubular cytoskeleton of mitotic cells using wild-type, mpk6-2 mutant and plants overexpressing the MAP kinase-inactivating phosphatase, AP2C3. We showed that MPK6 interacted with γ-tubulin and co-sedimented with plant microtubules polymerized in vitro. It was the active form of MAP kinase that was enriched with microtubules and followed similar dynamics to γ-tubulin, moving from poles to midzone during the anaphase-to-telophase transition. We found a novel substrate for MPK6, the microtubule plus end protein, EB1c. The mpk6-2 mutant was sensitive to 3-nitro-l-tyrosine (NO2-Tyr) treatment with respect to mitotic abnormalities, and root cells overexpressing AP2C3 showed defects in chromosome segregation and spindle orientation. Our data suggest that the active form of MAP kinase interacts with γ-tubulin on specific subsets of mitotic microtubules during late mitosis. MPK6 phosphorylates EB1c, but not EB1a, and has a role in maintaining regular planes of cell division under stress conditions.

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