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© The Rockefeller University Press, 0021-9525/1997//975 $5.00
The Journal of Cell Biology, Volume 139, Number 4, , 1997 975-983

Article

XMAP310: A Xenopus Rescue-promoting Factor Localized to the Mitotic Spindle



Søren S.L. Andersen and Eric Karsenti

European Molecular Biology Laboratory, Cell Biology Programme, D-69117 Heidelberg, Germany

To understand the role of microtubule-associated proteins (MAPs) in the regulation of microtubule (MT) dynamics we have characterized MAPs prepared from Xenopus laevis eggs (Andersen, S.S.L., B. Buendia, J.E. Domínguez, A. Sawyer, and E. Karsenti. 1994. J. Cell Biol. 127:1289–1299). Here we report on the purification and characterization of a 310-kD MAP (XMAP310) that localizes to the nucleus in interphase and to mitotic spindle MTs in mitosis. XMAP310 is present in eggs, oocytes, a Xenopus tissue culture cell line, testis, and brain. We have purified XMAP310 to homogeneity from egg extracts. The purified protein cross-links pure MTs. Analysis of the effect of this protein on MT dynamics by time-lapse video microscopy has shown that it increases the rescue frequency 5–10-fold and decreases the shrinkage rate twofold. It has no effect on the growth rate or the catastrophe frequency. Microsequencing data suggest that XMAP230 and XMAP310 are novel MAPs. Although the three Xenopus MAPs characterized so far, XMAP215 (Vasquez, R.J., D.L. Gard, and L. Cassimeris. 1994. J. Cell Biol. 127:985–993), XMAP230, and XMAP310 are localized to the mitotic spindle, they have distinct effects on MT dynamics. While XMAP215 promotes rapid MT growth, XMAP230 decreases the catastrophe frequency and XMAP310 increases the rescue frequency. This may have important implications for the regulation of MT dynamics during spindle morphogenesis and chromosome segregation.

1. Abbreviations used in this paper: fcat, catastrophe frequency; fres, rescue frequency; mAb, monoclonal antibody; MALDI MS, Matrix Assisted Laser Desorption Ionization Mass Spectroscopy; MAP, microtubule-associated protein; MT, microtubule; pAb, polyclonal antibody; vg, growth rate; vs, shrinkage rate.

Søren Andersen was partially supported by an individual European Union Marie Curie Grant (ERB-CHBIT 930755) and an EU network Grant to Tony Hyman; Eric Karsenti by a Human Frontier Science Program Grant (RG-350/94) as well as a European Union Grant (ERB-CHRXCT 940568).


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