A novel Crumbs3 isoform regulates cell division and ciliogenesis via importin beta interactions

J Cell Biol. 2007 Jul 30;178(3):387-98. doi: 10.1083/jcb.200609096. Epub 2007 Jul 23.

Abstract

The Crumbs family of apical transmembrane proteins regulates apicobasal polarity via protein interactions with a conserved C-terminal sequence, ERLI. However, one of the mammalian Crumbs proteins, Crumbs3 (CRB3) has an alternate splice form with a novel C-terminal sequence ending in CLPI (CRB3-CLPI). We report that CRB3-CLPI localizes to the cilia membrane and a membrane compartment at the mitotic spindle poles. Knockdown of CRB3-CLPI leads to both a loss of cilia and a multinuclear phenotype associated with centrosomal and spindle abnormalities. Using protein purification, we find that CRB3-CLPI interacts with importin beta-1 in a Ran-regulated fashion. Importin beta-1 colocalizes with CRB3-CLPI during mitosis, and a dominant-negative form of importin beta-1 closely phenocopies CRB3-CLPI knockdown. Knockdown of importin beta-1 blocks targeting of CRB3-CLPI to the spindle poles. Our data suggest an expanded role for Crumbs proteins in polarized membrane targeting and cell division via unique interactions with importin proteins.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • Alternative Splicing
  • Amino Acid Sequence
  • Animals
  • Cell Division / physiology*
  • Cell Line
  • Cell Membrane / metabolism
  • Cell Membrane / ultrastructure
  • Cell Polarity
  • Centrosome / metabolism
  • Centrosome / ultrastructure
  • Cilia / physiology*
  • Cilia / ultrastructure
  • Gene Targeting
  • Humans
  • Membrane Glycoproteins / genetics
  • Membrane Glycoproteins / metabolism*
  • Molecular Sequence Data
  • Protein Isoforms / genetics
  • Protein Isoforms / metabolism*
  • Sequence Alignment
  • Spindle Apparatus / metabolism
  • Spindle Apparatus / ultrastructure
  • beta Karyopherins / genetics
  • beta Karyopherins / metabolism*

Substances

  • CRB3 protein, human
  • KPNB1 protein, human
  • Membrane Glycoproteins
  • Protein Isoforms
  • beta Karyopherins