title: Coordination of adjacent domains mediates TACC3-ch-TOG-clathrin assembly and mitotic spindle binding.
abstract: A complex of transforming acidic coiled-coil protein 3 (TACC3), colonic and hepatic tumor overexpressed gene (ch-TOG), and clathrin has been implicated in mitotic spindle assembly and in the stabilization of kinetochore fibers by cross-linking microtubules. It is unclear how this complex binds microtubules and how the proteins in the complex interact with one another. TACC3 and clathrin have each been proposed to be the spindle recruitment factor. We have mapped the interactions within the complex and show that TACC3 and clathrin were interdependent for spindle recruitment, having to interact in order for either to be recruited to the spindle. The N-terminal domain of clathrin and the TACC domain of TACC3 in tandem made a microtubule interaction surface, coordinated by TACC3-clathrin binding. A dileucine motif and Aurora A-phosphorylated serine 558 on TACC3 bound to the "ankle" of clathrin. The other interaction within the complex involved a stutter in the TACC3 coiled-coil and a proposed novel sixth TOG domain in ch-TOG, which was required for microtubule localization of ch-TOG but not TACC3-clathrin.
authors.affiliation: Department of Cellular and Molecular Physiology, University of Liverpool, Liverpool L69 3BX, England, UK.
authors.forename: Fiona E
authors.initials: FE
authors.lastname: Hood
authors.forename: Samantha J
authors.initials: SJ
authors.lastname: Williams
authors.forename: Selena G
authors.initials: SG
authors.lastname: Burgess
authors.forename: Mark W
authors.initials: MW
authors.lastname: Richards
authors.forename: Daniel
authors.initials: D
authors.lastname: Roth
authors.forename: Anne
authors.initials: A
authors.lastname: Straube
authors.forename: Mark
authors.initials: M
authors.lastname: Pfuhl
authors.forename: Richard
authors.initials: R
authors.lastname: Bayliss
authors.forename: Stephen J
authors.initials: SJ
authors.lastname: Royle
mesh_terms: D000818:Animals; D001665:Binding Sites; D002352:Carrier Proteins / Q000378:metabolism*; D002966:Clathrin / Q000378:metabolism*; D005326:Fetal Proteins / Q000378:metabolism*; D057809:HEK293 Cells; D006801:Humans; D051379:Mice; D008869:Microtubule-Associated Proteins / Q000378:metabolism*; D008941:Spindle Apparatus / Q000378:metabolism*
chemical_list: D002352:Carrier Proteins; D002966:Clathrin; D005326:Fetal Proteins; D008869:Microtubule-Associated Proteins; C418485:TACC3 protein, mouse
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