Xenopus TACC3/maskin is not required for microtubule stability but is required for anchoring microtubules at the centrosome.
Members of the transforming acidic coiled coil (TACC) protein family are emerging as important mitotic spindle assembly proteins in a variety of organisms. The molecular details of how TACC proteins function are unknown, but TACC proteins have been proposed to recruit microtubule-stabilizing proteins of the tumor overexpressed gene (TOG) family to the centrosome and to facilitate their loading onto newly emerging microtubules. Using Xenopus egg extracts and in vitro assays, we show that the Xenopus TACC protein maskin is required for centrosome function beyond recruiting the Xenopus TOG protein XMAP215. The conserved C-terminal TACC domain of maskin is both necessary and sufficient to restore centrosome function in maskin-depleted extracts, and we provide evidence that the N terminus of maskin inhibits the function of the TACC domain. Time-lapse video microscopy reveals that microtubule dynamics in Xenopus egg extracts are unaffected by maskin depletion. Our results provide direct experimental evidence of a role for maskin in centrosome function and suggest that maskin is required for microtubule anchoring at the centrosome.
Albee
Alison J
AJ
Department of Biochemistry, University of Wisconsin-Madison, Madison, WI 53706, USA.
Wiese
Christiane
C
D000818:Animals; D018385:Centrosome / Q000378:metabolism*; D005260:Female; D008297:Male; D018715:Microscopy, Video; D008870:Microtubules / Q000737:chemistry / Q000378:metabolism*; D008954:Models, Biological; D017434:Protein Structure, Tertiary; D011994:Recombinant Proteins / Q000737:chemistry; D013094:Spermatozoa / Q000378:metabolism; D008941:Spindle Apparatus; D014157:Transcription Factors / Q000378:metabolism*; D014404:Tubulin / Q000378:metabolism; D029867:Xenopus Proteins / Q000378:metabolism*; D014982:Xenopus laevis
D011994:Recombinant Proteins; C474872:TACC3 protein, Xenopus; D014157:Transcription Factors; D014404:Tubulin; D029867:Xenopus Proteins