Involvement of PKC? and GSK3? in the stability of the metaphase spindle

Madhavi Kalive, Debra Baluch, David Capco

Research output: Contribution to journalArticle

Abstract

In the somatic cell, the mitotic spindle apparatus is centrosomal, and several isoforms of protein kinase C (PKC) have been associated with the mitotic spindle, but their role in stabilizing the mitotic spindle is still unclear. Other protein kinases such as, glycogen synthase kinase 3? (GSK3?) have also been shown to be associated with the mitotic spindle apparatus. In this study, we show the enrichment of active (phosphorylated) PKC? at the centrosomal region of the spindle apparatus in metaphase stage of 3T3 cells. In order to understand whether the two kinases PKC and GSK3? are associated with the mitotic spindle, first, the co-localization of phosphorylated PKC isoforms with GSK3? was studied at the poles in metaphase cells. Fluorescence resonance energy transfer (FRET) analysis was used to demonstrate close molecular proximity of phospho-PKC? with phospho(ser9)GSK3?. Second, the involvement of inactive GSK3? in maintaining an intact mitotic spindle in 3T3 cells was shown. Third, this study also showed that addition of a phospho-PKC? specific inhibitor to cells can disrupt the mitotic spindle microtubules and some of the proteins associated with it. The mitotic spindle at metaphase in mouse fibroblasts appears to be maintained by PKC? acting through GSK3?. Phospho-PKC? is in close molecular proximity to GSK3?, whereas the other isoforms of PKC such as pPKC?II, pPKC?, pPKC?, and pPKC? are not close enough to have significant FRET readings. The close molecular proximity supports the idea that GSK3? may be a substrate of PKC?.

Original languageEnglish (US)
Pages (from-to)97-111
Number of pages15
JournalIn Vitro Cellular and Developmental Biology - Animal
Volume48
Issue number2
DOIs
StatePublished - Feb 2012

Fingerprint

Spindle Apparatus
Metaphase
Protein Kinase C
3T3 Cells
Fluorescence Resonance Energy Transfer
Protein Isoforms
Glycogen Synthase Kinase 3
Microtubule-Associated Proteins
Protein Kinases
Reading
Fibroblasts

Keywords

  • Centromeres
  • FRET. Permeabilized cell
  • GSK
  • Kinases
  • Mitotic spindle
  • PKC

ASJC Scopus subject areas

  • Developmental Biology
  • Cell Biology
  • Medicine(all)

Cite this

Involvement of PKC? and GSK3? in the stability of the metaphase spindle. / Kalive, Madhavi; Baluch, Debra; Capco, David.

In: In Vitro Cellular and Developmental Biology - Animal, Vol. 48, No. 2, 02.2012, p. 97-111.

Research output: Contribution to journalArticle

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