H-ABC- and Dystonia-causing Mutations Show Distinct Pathogenic Effects

Overview

Journal Sci Adv

Specialties Biology
Science

Date 2022 Mar 11

PMID 35275727

Authors

Victor Krajka

Franca Vulinovic

Mariya Genova

Kerstin Tanzer

A S Jijumon

Satish Bodakuntla

Stephanie Tennstedt

Helge Mueller-Fielitz

Britta Meier

Carsten Janke

Christine Klein

Aleksandar Rakovic

Affiliations

Soon will be listed here.

Abstract

Mutations in the brain-specific β-tubulin 4A (TUBB4A) gene cause a broad spectrum of diseases, ranging from dystonia (DYT-TUBB4A) to hypomyelination with atrophy of the basal ganglia and cerebellum (H-ABC). Currently, the mechanisms of how variants lead to this pleiotropic manifestation remain elusive. Here, we investigated whether mutations causing either DYT-TUBB4A (p.R2G and p.Q424H) or H-ABC (p.R2W and p.D249N) exhibit differential effects at the molecular and cellular levels. Using live-cell imaging of disease-relevant oligodendrocytes and total internal reflection fluorescence microscopy of whole-cell lysates, we observed divergent impact on microtubule polymerization and microtubule integration, partially reflecting the observed pleiotropy. Moreover, in silico simulations demonstrated that the mutants rarely adopted a straight heterodimer conformation in contrast to wild type. In conclusion, for most of the examined variants, we deciphered potential molecular disease mechanisms that may lead to the diverse clinical manifestations and phenotype severity across and within each TUBB4A-related disease.

Citing Articles

Identification of a functional missense variant in the matrix metallopeptidase 10 (MMP10) gene in two families with premature myocardial infarction.

Verovenko V, Tennstedt S, Kleinecke M, Kessler T, Schunkert H, Erdmann J Sci Rep. 2024; 14(1):12212.

PMID: 38806571 PMC: 11133425. DOI: 10.1038/s41598-024-62878-3.

Deep brain stimulation in an adolescent with hypomyelination with atrophy of the basal ganglia and cerebellum due to a TUBB4A mutation: illustrative case.

MacLean J, Nataraj J, Olaya J, Liker M, Sanger T J Neurosurg Case Lessons. 2023; 6(2).

PMID: 37458337 PMC: 10555642. DOI: 10.3171/CASE23158.

The molecular biology of tubulinopathies: Understanding the impact of variants on tubulin structure and microtubule regulation.

Hoff K, Neumann A, Moore J Front Cell Neurosci. 2022; 16:1023267.

PMID: 36406756 PMC: 9666403. DOI: 10.3389/fncel.2022.1023267.

The organization of microtubules and Tau in oligodendrocytes: Tau pathology in damaged oligodendrocytes.

Torii T, Miyasaka T, Misonou H Front Cell Dev Biol. 2022; 10:950682.

PMID: 36274848 PMC: 9579343. DOI: 10.3389/fcell.2022.950682.

Understanding molecular mechanisms and predicting phenotypic effects of pathogenic tubulin mutations.

Attard T, Welburn J, Marsh J PLoS Comput Biol. 2022; 18(10):e1010611.

PMID: 36206299 PMC: 9581425. DOI: 10.1371/journal.pcbi.1010611.

References

Rosewich H, Thiele H, Ohlenbusch A, Maschke U, Altmuller J, Frommolt P . Heterozygous de-novo mutations in ATP1A3 in patients with alternating hemiplegia of childhood: a whole-exome sequencing gene-identification study. Lancet Neurol. 2012; 11(9):764-73. DOI: 10.1016/S1474-4422(12)70182-5. View

Sastry G, Adzhigirey M, Day T, Annabhimoju R, Sherman W . Protein and ligand preparation: parameters, protocols, and influence on virtual screening enrichments. J Comput Aided Mol Des. 2013; 27(3):221-34. DOI: 10.1007/s10822-013-9644-8. View

Dutta P, Jijumon A, Mazumder M, Dileep D, Mukhopadhyay A, Gourinath S . Presence of actin binding motif in VgrG-1 toxin of Vibrio cholerae reveals the molecular mechanism of actin cross-linking. Int J Biol Macromol. 2019; 133:775-785. DOI: 10.1016/j.ijbiomac.2019.04.026. View

Zhang Y, Sloan S, Clarke L, Caneda C, Plaza C, Blumenthal P . Purification and Characterization of Progenitor and Mature Human Astrocytes Reveals Transcriptional and Functional Differences with Mouse. Neuron. 2015; 89(1):37-53. PMC: 4707064. DOI: 10.1016/j.neuron.2015.11.013. View

Hersheson J, Mencacci N, Davis M, MacDonald N, Trabzuni D, Ryten M . Mutations in the autoregulatory domain of β-tubulin 4a cause hereditary dystonia. Ann Neurol. 2013; 73(4):546-53. PMC: 3698699. DOI: 10.1002/ana.23832. View

Douvaras P, Fossati V . Generation and isolation of oligodendrocyte progenitor cells from human pluripotent stem cells. Nat Protoc. 2015; 10(8):1143-54. DOI: 10.1038/nprot.2015.075. View

Carvalho D, Santos S, Martins B, Marques F . TUBB4A novel mutation reinforces the genotype-phenotype correlation of hypomyelination with atrophy of the basal ganglia and cerebellum. Brain. 2014; 138(Pt 2):e327. DOI: 10.1093/brain/awu242. View

Li W, Moriwaki T, Tani T, Watanabe T, Kaibuchi K, Goshima G . Reconstitution of dynamic microtubules with Drosophila XMAP215, EB1, and Sentin. J Cell Biol. 2012; 199(5):849-62. PMC: 3514792. DOI: 10.1083/jcb.201206101. View

Jaqaman K, Loerke D, Mettlen M, Kuwata H, Grinstein S, Schmid S . Robust single-particle tracking in live-cell time-lapse sequences. Nat Methods. 2008; 5(8):695-702. PMC: 2747604. DOI: 10.1038/nmeth.1237. View

10.

Aiken J, Buscaglia G, Bates E, Moore J . The -Tubulin gene in Brain Development: A Key Ingredient in the Neuronal Isotype Blend. J Dev Biol. 2017; 5(3). PMC: 5648057. DOI: 10.3390/jdb5030008. View

11.

Zheng C, Diaz-Cuadros M, Nguyen K, Hall D, Chalfie M . Distinct effects of tubulin isotype mutations on neurite growth in . Mol Biol Cell. 2017; 28(21):2786-2801. PMC: 5638583. DOI: 10.1091/mbc.E17-06-0424. View

12.

de Carvalho Aguiar P, Sweadner K, Penniston J, Zaremba J, Liu L, Caton M . Mutations in the Na+/K+ -ATPase alpha3 gene ATP1A3 are associated with rapid-onset dystonia parkinsonism. Neuron. 2004; 43(2):169-75. DOI: 10.1016/j.neuron.2004.06.028. View

13.

Yen T, Machlin P, Cleveland D . Autoregulated instability of beta-tubulin mRNAs by recognition of the nascent amino terminus of beta-tubulin. Nature. 1988; 334(6183):580-5. DOI: 10.1038/334580a0. View

14.

Gell C, Bormuth V, Brouhard G, Cohen D, Diez S, Friel C . Microtubule dynamics reconstituted in vitro and imaged by single-molecule fluorescence microscopy. Methods Cell Biol. 2010; 95:221-45. DOI: 10.1016/S0091-679X(10)95013-9. View

15.

Allen M, Poggiali D, Whitaker K, Marshall T, van Langen J, Kievit R . Raincloud plots: a multi-platform tool for robust data visualization. Wellcome Open Res. 2023; 4:63. PMC: 6480976. DOI: 10.12688/wellcomeopenres.15191.2. View

16.

Cao L, Wang W, Jiang Q, Wang C, Knossow M, Gigant B . The structure of apo-kinesin bound to tubulin links the nucleotide cycle to movement. Nat Commun. 2014; 5:5364. DOI: 10.1038/ncomms6364. View

17.

Zimmermann L, Stephens A, Nam S, Rau D, Kubler J, Lozajic M . A Completely Reimplemented MPI Bioinformatics Toolkit with a New HHpred Server at its Core. J Mol Biol. 2017; 430(15):2237-2243. DOI: 10.1016/j.jmb.2017.12.007. View

18.

Cushion T, Paciorkowski A, Pilz D, Mullins J, Seltzer L, Marion R . De novo mutations in the beta-tubulin gene TUBB2A cause simplified gyral patterning and infantile-onset epilepsy. Am J Hum Genet. 2014; 94(4):634-41. PMC: 3980418. DOI: 10.1016/j.ajhg.2014.03.009. View

19.

Castoldi M, Popov A . Purification of brain tubulin through two cycles of polymerization-depolymerization in a high-molarity buffer. Protein Expr Purif. 2003; 32(1):83-8. DOI: 10.1016/S1046-5928(03)00218-3. View

20.

Zhang Y, Chen K, Sloan S, Bennett M, Scholze A, OKeeffe S . An RNA-sequencing transcriptome and splicing database of glia, neurons, and vascular cells of the cerebral cortex. J Neurosci. 2014; 34(36):11929-47. PMC: 4152602. DOI: 10.1523/JNEUROSCI.1860-14.2014. View