Dephosphorylation of MAP2D enhances its binding to vimentin in preovulatory ovarian granulosa cells

Maxfield P. Flynn, Sarah E. Fiedler, Amelia B. Karlsson, Daniel Carr, Evelyn T. Maizels, Mary Hunzicker-Dunn

Research output: Contribution to journalArticle

3 Citations (Scopus)

Abstract

Preovulatory granulosa cells express the low-molecular-mass MAP2D variant of microtubule-associated protein 2 (MAP2). Activation of the luteinizing hormone choriogonadotropin receptor by human choriogonadotropin (hCG) promotes dephosphorylation of MAP2D on Thr256 and Thr259. We sought to evaluate the association of MAP2D with the cytoskeleton, and the effect of hCG on this association. MAP2D partially colocalized, as assessed by confocal immunofluorescence microscopy, with the vimentin intermediate filament and microtubule cytoskeletons in naive cells. In vitro binding studies showed that MAP2D bound directly to vimentin and β-tubulin. Phosphorylation of recombinantMAP2D on Thr256 and Thr259, which mimics the phosphorylation status of MAP2D in naive cells, reduces binding of MAP2D to vimentin and tubulin by two- and three-fold, respectively. PKA-dependent phosphorylation of vimentin (Ser32 and Ser38) promoted binding of vimentin to MAP2D and increased contraction of granulosa cells with reorganization of vimentin filaments and MAP2D from the periphery into a thickened layer surrounding the nucleus and into prominent cellular extensions. Chemical disruption of vimentin filament organization increased progesterone production. Taken together, these results suggest that hCG-stimulated dephosphorylation of MAP2D at Thr256 and Thr259, phosphorylation of vimentin at Ser38 and Ser72, and the resulting enhanced binding of MAP2D to vimentin might contribute to the progesterone synthetic response required for ovulation.

Original languageEnglish (US)
Pages (from-to)2983-2996
Number of pages14
JournalJournal of Cell Science
Volume129
Issue number15
DOIs
StatePublished - 2016

Fingerprint

Granulosa Cells
Vimentin
Chorionic Gonadotropin
Phosphorylation
Tubulin
Cytoskeleton
Progesterone
LH Receptors
Microtubule-Associated Proteins
Intermediate Filaments
Ovulation
Fluorescence Microscopy
Confocal Microscopy
Microtubules

Keywords

  • Choriogonadotropin receptor
  • Granulosa cell
  • Luteinizing hormone receptor
  • MAP2D
  • PKA
  • Vimentin

ASJC Scopus subject areas

  • Cell Biology

Cite this

Flynn, M. P., Fiedler, S. E., Karlsson, A. B., Carr, D., Maizels, E. T., & Hunzicker-Dunn, M. (2016). Dephosphorylation of MAP2D enhances its binding to vimentin in preovulatory ovarian granulosa cells. Journal of Cell Science, 129(15), 2983-2996. https://doi.org/10.1242/jcs.190397

Dephosphorylation of MAP2D enhances its binding to vimentin in preovulatory ovarian granulosa cells. / Flynn, Maxfield P.; Fiedler, Sarah E.; Karlsson, Amelia B.; Carr, Daniel; Maizels, Evelyn T.; Hunzicker-Dunn, Mary.

In: Journal of Cell Science, Vol. 129, No. 15, 2016, p. 2983-2996.

Research output: Contribution to journalArticle

Flynn, MP, Fiedler, SE, Karlsson, AB, Carr, D, Maizels, ET & Hunzicker-Dunn, M 2016, 'Dephosphorylation of MAP2D enhances its binding to vimentin in preovulatory ovarian granulosa cells', Journal of Cell Science, vol. 129, no. 15, pp. 2983-2996. https://doi.org/10.1242/jcs.190397
Flynn, Maxfield P. ; Fiedler, Sarah E. ; Karlsson, Amelia B. ; Carr, Daniel ; Maizels, Evelyn T. ; Hunzicker-Dunn, Mary. / Dephosphorylation of MAP2D enhances its binding to vimentin in preovulatory ovarian granulosa cells. In: Journal of Cell Science. 2016 ; Vol. 129, No. 15. pp. 2983-2996.
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