Drosophila EB1 is important for proper assembly, dynamics, and positioning of the mitotic spindle

Stephen L. Rogers, Gregory C. Rogers, David J. Sharp, Ronald D. Vale

Research output: Contribution to journalArticle

291 Citations (Scopus)

Abstract

EB1 is an evolutionarily conserved protein that localizes to the plus ends of growing microtubules. In yeast, the EB1 homologue (BIMI) has been shown to modulate microtubule dynamics and link microtubules to the cortex, but the functions of metazoan EB1 proteins remain unknown. Using a novel preparation of the Drosophila S2 cell line that promotes cell attachment and spreading, we visualized dynamics of single microtubules in real time and found that depletion of EB1 by RNA-mediated inhibition (RNAi) in interphase cells causes a dramatic increase in nondynamic microtubules (neither growing nor shrinking), but does not alter overall microtubule organization. In contrast, several defects in microtubule organization are observed in RNAi-treated mitotic cells, including a drastic reduction in astral microtubules, malformed mitotic spindles, defocused spindle poles, and mispositioning of spindles away from the cell center. Similar phenotypes were observed in mitotic spindles of Drosophila embryos that were microinjected with anti-EB1 antibodies. In addition, live cell imaging of mitosis in Drosophila embryos reveals defective spindle elongation and chromosomal segregation during anaphase after antibody injection. Our results reveal crucial roles for EB1 in mitosis, which we postulate involves its ability to promote the growth and interactions of microtubules within the central spindle and at the cell cortex.

Original languageEnglish (US)
Pages (from-to)873-884
Number of pages12
JournalJournal of Cell Biology
Volume158
Issue number5
DOIs
StatePublished - Sep 2 2002

Fingerprint

Spindle Apparatus
Microtubules
Drosophila
Mitosis
Embryonic Structures
Spindle Poles
RNA
Anaphase
Interphase
Anti-Idiotypic Antibodies
Proteins
Yeasts
Phenotype
Cell Line
Injections
Antibodies

Keywords

  • Dynamics
  • EB1
  • Microtubule
  • Mitosis
  • Spindle

ASJC Scopus subject areas

  • Cell Biology

Cite this

Drosophila EB1 is important for proper assembly, dynamics, and positioning of the mitotic spindle. / Rogers, Stephen L.; Rogers, Gregory C.; Sharp, David J.; Vale, Ronald D.

In: Journal of Cell Biology, Vol. 158, No. 5, 02.09.2002, p. 873-884.

Research output: Contribution to journalArticle

Rogers, Stephen L. ; Rogers, Gregory C. ; Sharp, David J. ; Vale, Ronald D. / Drosophila EB1 is important for proper assembly, dynamics, and positioning of the mitotic spindle. In: Journal of Cell Biology. 2002 ; Vol. 158, No. 5. pp. 873-884.
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