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Mol Biol Cell
2002 Oct 01;1310:3614-26. doi: 10.1091/mbc.02-04-0210.
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EB1-microtubule interactions in Xenopus egg extracts: role of EB1 in microtubule stabilization and mechanisms of targeting to microtubules.
Tirnauer JS
,
Grego S
,
Salmon ED
,
Mitchison TJ
.
???displayArticle.abstract??? EB1 targets to polymerizing microtubule ends, where it is favorably positioned to regulate microtubule polymerization and confer molecular recognition of the microtubule end. In this study, we focus on two aspects of the EB1-microtubule interaction: regulation of microtubule dynamics by EB1 and the mechanism of EB1 association with microtubules. Immunodepletion of EB1 from cytostatic factor-arrested M-phase Xenopus egg extracts dramatically reduced microtubule length; this was complemented by readdition of EB1. By time-lapse microscopy, EB1 increased the frequency of microtubule rescues and decreased catastrophes, resulting in increased polymerization and decreased depolymerization and pausing. Imaging of EB1 fluorescence revealed a novel structure: filamentous extensions on microtubule plus ends that appeared during microtubule pauses; loss of these extensions correlated with the abrupt onset of polymerization. Fluorescent EB1 localized to comets at the polymerizing plus ends of microtubules in cytostatic factor extracts and uniformly along the lengths of microtubules in interphase extracts. The temporal decay of EB1 fluorescence from polymerizing microtubule plus ends predicted a dissociation half-life of seconds. Fluorescence recovery after photobleaching also revealed dissociation and rebinding of EB1 to the microtubule wall with a similar half-life. EB1 targeting to microtubules is thus described by a combination of higher affinity binding to polymerizing ends and lower affinity binding along the wall, with continuous dissociation. The latter is likely to be attenuated in interphase. The highly conserved effect of EB1 on microtubule dynamics suggests it belongs to a core set of regulatory factors conserved in higher organisms, and the complex pattern of EB1 targeting to microtubules could be exploited by the cell for coordinating microtubule behaviors.
Adames,
Microtubule interactions with the cell cortex causing nuclear movements in Saccharomyces cerevisiae.
2000, Pubmed
Adames,
Microtubule interactions with the cell cortex causing nuclear movements in Saccharomyces cerevisiae.
2000,
Pubmed
Akhmanova,
Clasps are CLIP-115 and -170 associating proteins involved in the regional regulation of microtubule dynamics in motile fibroblasts.
2001,
Pubmed
Arnal,
Structural transitions at microtubule ends correlate with their dynamic properties in Xenopus egg extracts.
2000,
Pubmed
,
Xenbase
Beinhauer,
Mal3, the fission yeast homologue of the human APC-interacting protein EB-1 is required for microtubule integrity and the maintenance of cell form.
1997,
Pubmed
Berrueta,
The adenomatous polyposis coli-binding protein EB1 is associated with cytoplasmic and spindle microtubules.
1998,
Pubmed
Bu,
Regulation of microtubule assembly by human EB1 family proteins.
2001,
Pubmed
Bulinski,
Rapid dynamics of the microtubule binding of ensconsin in vivo.
2001,
Pubmed
Cassimeris,
Regulation of microtubule-associated proteins.
2001,
Pubmed
Davis,
Purification and biochemical characterization of tubulin from the budding yeast Saccharomyces cerevisiae.
1993,
Pubmed
Desai,
Microtubule polymerization dynamics.
1997,
Pubmed
Desai,
Anaphase A chromosome movement and poleward spindle microtubule flux occur At similar rates in Xenopus extract spindles.
1998,
Pubmed
,
Xenbase
Desai,
The use of Xenopus egg extracts to study mitotic spindle assembly and function in vitro.
1999,
Pubmed
,
Xenbase
Desai,
Kin I kinesins are microtubule-destabilizing enzymes.
1999,
Pubmed
,
Xenbase
Diamantopoulos,
Dynamic localization of CLIP-170 to microtubule plus ends is coupled to microtubule assembly.
1999,
Pubmed
Fodde,
Mutations in the APC tumour suppressor gene cause chromosomal instability.
2001,
Pubmed
Gard,
Microtubule assembly in cytoplasmic extracts of Xenopus oocytes and eggs.
1987,
Pubmed
,
Xenbase
Grego,
Microtubule treadmilling in vitro investigated by fluorescence speckle and confocal microscopy.
2001,
Pubmed
Hyman,
Preparation of modified tubulins.
1991,
Pubmed
Hyman,
Role of GTP hydrolysis in microtubule dynamics: information from a slowly hydrolyzable analogue, GMPCPP.
1992,
Pubmed
Jánosi,
Modeling elastic properties of microtubule tips and walls.
1998,
Pubmed
Kaplan,
A role for the Adenomatous Polyposis Coli protein in chromosome segregation.
2001,
Pubmed
Kinoshita,
Reconstitution of physiological microtubule dynamics using purified components.
2001,
Pubmed
,
Xenbase
Korinek,
Molecular linkage underlying microtubule orientation toward cortical sites in yeast.
2000,
Pubmed
Lee,
Control of mitotic spindle position by the Saccharomyces cerevisiae formin Bni1p.
1999,
Pubmed
Maddox,
The polarity and dynamics of microtubule assembly in the budding yeast Saccharomyces cerevisiae.
2000,
Pubmed
Mandelkow,
Microtubule dynamics and microtubule caps: a time-resolved cryo-electron microscopy study.
1991,
Pubmed
Miller,
Bim1p/Yeb1p mediates the Kar9p-dependent cortical attachment of cytoplasmic microtubules.
2000,
Pubmed
Mimori-Kiyosue,
The dynamic behavior of the APC-binding protein EB1 on the distal ends of microtubules.
2000,
Pubmed
,
Xenbase
Mitchison,
Isolation of mammalian centrosomes.
1986,
Pubmed
Nakamura,
Critical role for the EB1 and APC interaction in the regulation of microtubule polymerization.
2001,
Pubmed
Olmsted,
Cell cycle-dependent changes in the dynamics of MAP 2 and MAP 4 in cultured cells.
1989,
Pubmed
Palazzo,
mDia mediates Rho-regulated formation and orientation of stable microtubules.
2001,
Pubmed
Parsons,
Microtubule assembly in clarified Xenopus egg extracts.
1997,
Pubmed
,
Xenbase
Rusan,
Cell cycle-dependent changes in microtubule dynamics in living cells expressing green fluorescent protein-alpha tubulin.
2001,
Pubmed
Schroer,
Microtubules don and doff their caps: dynamic attachments at plus and minus ends.
2001,
Pubmed
Schuyler,
Microtubule "plus-end-tracking proteins": The end is just the beginning.
2001,
Pubmed
Schuyler,
Search, capture and signal: games microtubules and centrosomes play.
2001,
Pubmed
Theriot,
The rate of actin-based motility of intracellular Listeria monocytogenes equals the rate of actin polymerization.
1992,
Pubmed
Tirnauer,
EB1 proteins regulate microtubule dynamics, cell polarity, and chromosome stability.
2000,
Pubmed
Tirnauer,
Yeast Bim1p promotes the G1-specific dynamics of microtubules.
1999,
Pubmed
Tirnauer,
EB1 targets to kinetochores with attached, polymerizing microtubules.
2002,
Pubmed
Tran,
How tubulin subunits are lost from the shortening ends of microtubules.
1997,
Pubmed
Wadsworth,
Regional regulation of microtubule dynamics in polarized, motile cells.
1999,
Pubmed
Walker,
Dynamic instability of individual microtubules analyzed by video light microscopy: rate constants and transition frequencies.
1988,
Pubmed
Waterman-Storer,
Fluorescent speckle microscopy of microtubules: how low can you go?
1999,
Pubmed
Waterman-Storer,
Fluorescent speckle microscopy, a method to visualize the dynamics of protein assemblies in living cells.
1998,
Pubmed
,
Xenbase
Zhai,
Kinetochore microtubule dynamics and the metaphase-anaphase transition.
1995,
Pubmed