title: The lissencephaly protein Lis1 is present in motile mammalian cilia and requires outer arm dynein for targeting to Chlamydomonas flagella.
abstract: Lissencephaly is a developmental brain disorder characterized by a smooth cerebral surface, thickened cortex and misplaced neurons. Classical lissencephaly is caused by mutations in LIS1, which encodes a WD-repeat protein involved in cytoplasmic dynein regulation, mitosis and nuclear migration. Several proteins required for nuclear migration in Aspergillus bind directly to Lis1, including NudC. Mammalian NudC is highly expressed in ciliated epithelia, and localizes to motile cilia in various tissues. Moreover, a NudC ortholog is upregulated upon deflagellation in Chlamydomonas. We found that mammalian Lis1 localizes to motile cilia in trachea and oviduct, but is absent from non-motile primary cilia. Furthermore, we cloned a gene encoding a Lis1-like protein (CrLis1) from Chlamydomonas. CrLis1 is a approximately 37 kDa protein that contains seven WD-repeat domains, similar to Lis1 proteins from other organisms. Immunoblotting using an anti-CrLis1 antibody revealed that this protein is present in the flagellum and is depleted from flagella of mutants with defective outer dynein arm assembly, including one strain that lacks only the alpha heavy chain/light chain 5 thioredoxin complex. Biochemical experiments confirmed that CrLis1 associates with outer dynein arm components and revealed that CrLis1 binds directly to rat NudC. Our results suggest that Lis1 and NudC are present in cilia and flagella and may regulate outer dynein arm activity.
authors.affiliation: Department of Molecular Biology, University of Copenhagen, Universitetsparken 13, DK-2100 Copenhagen OE, Denmark. LBPedersen@aki.ku.dk
authors.forename: Lotte B
authors.initials: LB
authors.lastname: Pedersen
authors.forename: Panteleimon
authors.initials: P
authors.lastname: Rompolas
authors.forename: Søren T
authors.initials: ST
authors.lastname: Christensen
authors.forename: Joel L
authors.initials: JL
authors.lastname: Rosenbaum
authors.forename: Stephen M
authors.initials: SM
authors.lastname: King
mesh_terms: D043203:1-Alkyl-2-acetylglycerophosphocholine Esterase / Q000235:genetics / Q000378:metabolism*; D000595:Amino Acid Sequence; D000818:Animals; D015152:Blotting, Northern; D015139:Blotting, Southern; D002696:Chlamydomonas / Q000235:genetics / Q000378:metabolism*; D002923:Cilia / Q000378:metabolism*; D004398:Dyneins / Q000235:genetics / Q000378:metabolism*; D004591:Electrophoresis, Polyacrylamide Gel; D005260:Female; D005407:Flagella / Q000378:metabolism*; D006801:Humans; D051379:Mice; D008856:Microscopy, Fluorescence; D008869:Microtubule-Associated Proteins / Q000235:genetics / Q000378:metabolism*; D008969:Molecular Sequence Data; D041681:NIH 3T3 Cells; D009687:Nuclear Proteins / Q000235:genetics / Q000378:metabolism; D010053:Ovary / Q000378:metabolism; D010057:Oviducts / Q000378:metabolism; D010802:Phylogeny; D011485:Protein Binding; D011506:Proteins / Q000235:genetics / Q000378:metabolism; D051381:Rats; D011994:Recombinant Proteins / Q000378:metabolism; D017386:Sequence Homology, Amino Acid; D014132:Trachea / Q000378:metabolism
chemical_list: D008869:Microtubule-Associated Proteins; D009687:Nuclear Proteins; C094004:Nudc protein, rat; D011506:Proteins; D011994:Recombinant Proteins; D043203:1-Alkyl-2-acetylglycerophosphocholine Esterase; C513190:Pafah1b1 protein, mouse; D004398:Dyneins