A proteomic survey of microtubule-associated proteins in a R402H TUBA1A mutant mouse

Autoři: Ines Leca aff001;  Alexander Phillips aff001;  Iris Hofer aff001;  Lukas Landler aff001;  Lyubov Ushakova aff001;  Thomas David Cushion aff001;  Gerhard Dürnberger aff001;  Karel Stejskal aff001;  Karl Mechtler aff001;  David Anthony Keays aff001;  Alexander William Phillips aff001
Působiště autorů: Research Institute of Molecular Pathology (IMP), Vienna Biocenter (VBC), Vienna, Austria aff001;  Institute of Zoology, University of Natural Resources and Life Sciences (BOKU), Vienna, Austria aff002;  Gregor Mendel Institute (GMI), Austrian Academy of Sciences, Vienna Biocenter (VBC), Vienna, Austria aff003;  Institute of Molecular Biotechnology of the Austrian Academy of Sciences (IMBA), Vienna Biocenter (VBC), Vienna, Austria aff004;  Department of Anatomy and Neuroscience, University of Melbourne, Parkville, Australia aff005;  Division of Neurobiology, Department Biology II, Ludwig-Maximilians-University Munich, Planegg-Martinsried 82152, Germany aff006
Vyšlo v časopise: A proteomic survey of microtubule-associated proteins in a R402H TUBA1A mutant mouse. PLoS Genet 16(11): e32767. doi:10.1371/journal.pgen.1009104
Kategorie: Research Article
doi: 10.1371/journal.pgen.1009104


Microtubules play a critical role in multiple aspects of neurodevelopment, including the generation, migration and differentiation of neurons. A recurrent mutation (R402H) in the α-tubulin gene TUBA1A is known to cause lissencephaly with cerebellar and striatal phenotypes. Previous work has shown that this mutation does not perturb the chaperone-mediated folding of tubulin heterodimers, which are able to assemble and incorporate into the microtubule lattice. To explore the molecular mechanisms that cause the disease state we generated a new conditional mouse line that recapitulates the R402H variant. We show that heterozygous mutants present with laminar phenotypes in the cortex and hippocampus, as well as a reduction in striatal size and cerebellar abnormalities. We demonstrate that homozygous expression of the R402H allele causes neuronal death and exacerbates a cell intrinsic defect in cortical neuronal migration. Microtubule sedimentation assays coupled with quantitative mass spectrometry demonstrated that the binding and/or levels of multiple microtubule associated proteins (MAPs) are perturbed by the R402H mutation including VAPB, REEP1, EZRIN, PRNP and DYNC1l1/2. Consistent with these data we show that the R402H mutation impairs dynein-mediated transport which is associated with a decoupling of the nucleus to the microtubule organising center. Our data support a model whereby the R402H variant is able to fold and incorporate into microtubules, but acts as a gain of function by perturbing the binding of MAPs.

Klíčová slova:

Hippocampus – Homozygosity – Lysosomes – Microtubules – Motor proteins – Neurons – Tubulins – Neuron migration


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