Multiple mechanisms regulate NuMA dynamics at spindle poles

Olga Kisurina-Evgenieva, Gary Mack, Quansheng Du, Ian Macara, Alexey Khodjakov, Duane A. Compton

Research output: Contribution to journalArticle

44 Citations (Scopus)

Abstract

The large coiled-coil protein NuMA plays an essential role in organizing microtubule minus ends at spindle poles in vertebrate cells. Here, we use both in vivo and in vitro methods to examine NuMA dynamics at mitotic spindle poles. Using fluorescence recovery after photobleaching, we show that an exogenously expressed green-fluorescent-protein/NuMA fusion undergoes continuous exchange between soluble and spindle-associated pools in living cells. These dynamics require cellular energy and display an average half-time for fluorescence recovery of ∼3 minutes. To explore how NuMA dynamics at spindle poles is regulated, we exploited the association of NuMA with microtubule asters formed in mammalian mitotic extracts. Using a monoclonal antibody specific for human NuMA, we followed the fate of human NuMA associated with microtubule asters upon dilution with a hamster mitotic extract. Consistent with in vivo data, this assay shows that NuMA can be displaced from the core of pre-assembled asters into the soluble pool. The half-time of NuMA displacement from asters under these conditions is ∼5 minutes. Using this assay, we show that protein kinase activity and the NuMA-binding protein LGN regulate the dynamic exchange of NuMA on microtubule asters. Thus, the dynamic properties of NuMA are regulated by multiple mechanisms including protein phosphorylation and binding to the LGN protein, and the rate of exchange between soluble and microtubule-associated pools suggests that NuMA associates with an insoluble matrix at spindle poles.

Original languageEnglish (US)
Pages (from-to)6391-6400
Number of pages10
JournalJournal of Cell Science
Volume117
Issue number26
DOIs
StatePublished - Dec 15 2004

Fingerprint

Spindle Poles
Microtubules
Fluorescence Recovery After Photobleaching
Spindle Apparatus
Green Fluorescent Proteins
Protein Binding
Cricetinae
Protein Kinases
Vertebrates
Carrier Proteins
Proteins
Fluorescence
Monoclonal Antibodies
Phosphorylation

Keywords

  • Centrosome
  • Kinetochore
  • Microtubule
  • Mitotic spindle
  • NuMA

ASJC Scopus subject areas

  • Cell Biology

Cite this

Kisurina-Evgenieva, O., Mack, G., Du, Q., Macara, I., Khodjakov, A., & Compton, D. A. (2004). Multiple mechanisms regulate NuMA dynamics at spindle poles. Journal of Cell Science, 117(26), 6391-6400. https://doi.org/10.1242/jcs.01568

Multiple mechanisms regulate NuMA dynamics at spindle poles. / Kisurina-Evgenieva, Olga; Mack, Gary; Du, Quansheng; Macara, Ian; Khodjakov, Alexey; Compton, Duane A.

In: Journal of Cell Science, Vol. 117, No. 26, 15.12.2004, p. 6391-6400.

Research output: Contribution to journalArticle

Kisurina-Evgenieva, O, Mack, G, Du, Q, Macara, I, Khodjakov, A & Compton, DA 2004, 'Multiple mechanisms regulate NuMA dynamics at spindle poles', Journal of Cell Science, vol. 117, no. 26, pp. 6391-6400. https://doi.org/10.1242/jcs.01568
Kisurina-Evgenieva O, Mack G, Du Q, Macara I, Khodjakov A, Compton DA. Multiple mechanisms regulate NuMA dynamics at spindle poles. Journal of Cell Science. 2004 Dec 15;117(26):6391-6400. https://doi.org/10.1242/jcs.01568
Kisurina-Evgenieva, Olga ; Mack, Gary ; Du, Quansheng ; Macara, Ian ; Khodjakov, Alexey ; Compton, Duane A. / Multiple mechanisms regulate NuMA dynamics at spindle poles. In: Journal of Cell Science. 2004 ; Vol. 117, No. 26. pp. 6391-6400.
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