Disruption of a Ciliary B9 Protein Complex Causes Meckel Syndrome
Por:
W. DOWDLE, J. ROBINSON, A. KNEIST, M. SIREROL-PIQUER, S. FRINTS, K. CORBIT, N. ZAGHLOUL, G. VAN LIJNSCHOTEN, L. MULDERS, D. VERVER, K. ZERRES, R. REED, T. ATTIE-BITACH, C. JOHNSON, J. GARCIA-VERDUGO, N. KATSANIS, C. BERGMANN and J. REITER
Publicada:
15 jul 2011
Resumen:
Nearly every ciliated organism possesses three B9 domain-containing proteins: MKS1, B9D1, and B9D2. Mutations in human MKS1 cause Meckel syndrome (MKS), a severe ciliopathy characterized by occipital encephalocele, liver ductal plate malformations, polydactyly, and kidney cysts. Mouse mutations in either Mks1 or B9d2 compromise ciliogenesis and result in phenotypes similar to those of MKS. Given the importance of these two B9 proteins to ciliogenesis, we examined the role of the third B9 protein, B9d1. Mice lacking B9d1 displayed polydactyly, kidney cysts, ductal plate malformations, and abnormal patterning of the neural tube, concomitant with compromised ciliogenesis, ciliary protein localization, and Hedgehog (Hh) signal transduction. These data prompted us to screen MKS patients for mutations in B9D1 and B9D2. We identified a homozygous c.301A>C (p.Ser101Arg) B9D2 mutation that segregates with MKS, affects an evolutionarily conserved residue, and is absent from controls. Unlike wild-type B9D2 mRNA, the p.Ser101Arg mutation failed to rescue zebrafish phenotypes induced by the suppression of b9d2. With coimmunoprecipitation and mass spectrometric analyses, we found that Mks1, B9d1, and B9d2 interact physically, but that the p.Ser101Arg mutation abrogates the ability of B9d2 to interact with Mks1, further suggesting that the mutation compromises B9d2 function. Our data indicate that B9d1 is required for normal Hh signaling, ciliogenesis, and ciliary protein localization and that B9d1 and B9d2 are essential components of a B9 protein complex, disruption of which causes MKS.
Filiaciones:
W. DOWDLE:
Univ Calif San Francisco, Cardiovasc Res Inst, San Francisco, CA 94158 USA
Univ Calif San Francisco, Dept Biochem & Biophys, San Francisco, CA 94158 USA
J. ROBINSON:
Duke Univ, Dept Cell Biol, Ctr Human Dis Modeling, Durham, NC 27710 USA
Duke Univ, Dept Pediat, Ctr Human Dis Modeling, Durham, NC 27710 USA
A. KNEIST:
Rhein Westfal TH Aachen, Dept Human Genet, D-52056 Aachen, Germany
M. SIREROL-PIQUER:
CIBERNED, Unidad Mixta CIPF UVEG, Dept Cellular Morphol, Valencia 46012, Spain
S. FRINTS:
Maastricht Univ, Med Ctr, Dept Clin Genet Prenatal Diag & Therapy, NL-6200 MD Maastricht, Netherlands
Maastricht Univ, GROW, Res Sch Oncol & Dev Biol, NL-6200 MD Maastricht, Netherlands
K. CORBIT:
Univ Calif San Francisco, Cardiovasc Res Inst, San Francisco, CA 94158 USA
Univ Calif San Francisco, Dept Biochem & Biophys, San Francisco, CA 94158 USA
N. ZAGHLOUL:
Duke Univ, Dept Cell Biol, Ctr Human Dis Modeling, Durham, NC 27710 USA
Duke Univ, Dept Pediat, Ctr Human Dis Modeling, Durham, NC 27710 USA
Univ Maryland, Sch Med, Baltimore, MD 21201 USA
G. VAN LIJNSCHOTEN:
PAMM, Dept Pathol, NL-5623 EJ Eindhoven, Netherlands
L. MULDERS:
Maxima Med Ctr, Dept Gynecol, NL-5500 MB Veldhoven, Netherlands
D. VERVER:
Maastricht Univ, GROW, Res Sch Oncol & Dev Biol, NL-6200 MD Maastricht, Netherlands
Univ Paris 06, Dev Biol Lab, CNRS, UMR 7622, F-75005 Paris, France
K. ZERRES:
Rhein Westfal TH Aachen, Dept Human Genet, D-52056 Aachen, Germany
R. REED:
Johns Hopkins Sch Med, Ctr Sensory Biol, Baltimore, MD 21205 USA
T. ATTIE-BITACH:
Univ Paris 05, Hop Necker Enfants Malad, Dept Genet, F-75006 Paris, France
Univ Paris 05, Hop Necker Enfants Malad, INSERM, U781, F-75006 Paris, France
C. JOHNSON:
St James Univ Hosp, Leeds Inst Mol Med, Sect Ophthalmol & Neurosci, Leeds LS9 7TF, W Yorkshire, England
:
Univ Paris 06, Dev Biol Lab, CNRS, UMR 7622, F-75005 Paris, France
Cavanilles Inst Biodivers & Evolut Biol CIBERNED, Dept Comparat Neurol, Valencia 46980, Spain
N. KATSANIS:
Duke Univ, Dept Cell Biol, Ctr Human Dis Modeling, Durham, NC 27710 USA
Duke Univ, Dept Pediat, Ctr Human Dis Modeling, Durham, NC 27710 USA
C. BERGMANN:
Rhein Westfal TH Aachen, Dept Human Genet, D-52056 Aachen, Germany
Bioscientia, Ctr Human Genet, D-55218 Ingelheim, Germany
J. REITER:
Univ Calif San Francisco, Cardiovasc Res Inst, San Francisco, CA 94158 USA
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