title: The novel PAR-1-binding protein MTCL1 has crucial roles in organizing microtubules in polarizing epithelial cells.
abstract: The establishment of epithelial polarity is tightly linked to the dramatic reorganization of microtubules (MTs) from a radial array to a vertical alignment of non-centrosomal MT bundles along the lateral membrane, and a meshwork under the apical and basal membranes. However, little is known about the underlying molecular mechanism of this polarity-dependent MT remodeling. The evolutionarily conserved cell polarity-regulating kinase PAR-1 (known as MARK in mammals), whose activity is essential for maintaining the dynamic state of MTs, has indispensable roles in promoting this process. Here, we identify a novel PAR-1-binding protein, which we call microtubule crosslinking factor 1 (MTCL1), that crosslinks MTs through its N-terminal MT-binding region and subsequent coiled-coil motifs. MTCL1 colocalized with the apicobasal MT bundles in epithelial cells, and its knockdown impaired the development of these MT bundles and the epithelial-cell-specific columnar shape. Rescue experiments revealed that the N-terminal MT-binding region was indispensable for restoring these defects of the knockdown cells. MT regrowth assays indicated that MTCL1 was not required for the initial radial growth of MTs from the apical centrosome but was essential for the accumulation of non-centrosomal MTs to the sublateral regions. Interestingly, MTCL1 recruited a subpopulation of PAR-1b (known as MARK2 in mammals) to the apicobasal MT bundles, and its interaction with PAR-1b was required for MTCL1-dependent development of the apicobasal MT bundles. These results suggest that MTCL1 mediates the epithelial-cell-specific reorganization of non-centrosomal MTs through its MT-crosslinking activity, and cooperates with PAR-1b to maintain the correct temporal balance between dynamic and stable MTs within the apicobasal MT bundles.
authors.affiliation: Molecular Cellular Biology Laboratory, Yokohama City University Graduate School of Medical Life Science, 1-7-29 Suehirocho, Tsurumi, Yokohama 230-0045, Japan.
authors.forename: Yoshinori
authors.initials: Y
authors.lastname: Sato
authors.forename: Masashi
authors.initials: M
authors.lastname: Akitsu
authors.forename: Yoshiko
authors.initials: Y
authors.lastname: Amano
authors.forename: Kazunari
authors.initials: K
authors.lastname: Yamashita
authors.forename: Mariko
authors.initials: M
authors.lastname: Ide
authors.forename: Kyoko
authors.initials: K
authors.lastname: Shimada
authors.forename: Akio
authors.initials: A
authors.lastname: Yamashita
authors.forename: Hisashi
authors.initials: H
authors.lastname: Hirano
authors.forename: Noriaki
authors.initials: N
authors.lastname: Arakawa
authors.forename: Takahisa
authors.initials: T
authors.lastname: Maki
authors.forename: Ikuko
authors.initials: I
authors.lastname: Hayashi
authors.forename: Shigeo
authors.initials: S
authors.lastname: Ohno
authors.forename: Atsushi
authors.initials: A
authors.lastname: Suzuki
mesh_terms: D000818:Animals; D016764:Cell Polarity / Q000502:physiology; D002478:Cells, Cultured; D004285:Dogs; D004847:Epithelial Cells / Q000166:cytology* / Q000201:enzymology / Q000378:metabolism*; D057809:HEK293 Cells; D006367:HeLa Cells; D006801:Humans; D047468:Immunoprecipitation; D061985:Madin Darby Canine Kidney Cells; D051379:Mice; D008869:Microtubule-Associated Proteins / Q000378:metabolism*; D008870:Microtubules / Q000378:metabolism*; D009024:Morphogenesis; D011485:Protein Binding; D017346:Protein Serine-Threonine Kinases / Q000378:metabolism*; D014162:Transfection
chemical_list: C000604105:MTCL1 protein, human; D008869:Microtubule-Associated Proteins; D017346:Protein Serine-Threonine Kinases
keywords: Cell polarity; Epithelial cells; KIAA0802; MARK; MTCL1; Non-centrosomal microtubules; PAR-1