[en] Autosomal recessive cutis laxa type 2 (ARCL2), a syndrome of growth and developmental delay and redundant, inelastic skin, is caused by mutations in the a2 subunit of the vesicular ATPase H+-pump (ATP6V0A2). The goal of this study was to define the disease mechanisms that lead to connective tissue lesions in ARCL2. In a new cohort of 17 patients, DNA sequencing of ATP6V0A2 detected either homozygous or compound heterozygous mutations. Considerable allelic and phenotypic heterogeneity was observed, with a missense mutation of a moderately conserved residue p.P87L leading to unusually mild disease. Abnormal N- and/or mucin type O-glycosylation was observed in all patients tested. Premature stop codon mutations led to decreased ATP6V0A2 mRNA levels by destabilizing the mutant mRNA via the nonsense-mediated decay pathway. Loss of ATP6V0A2 either by siRNA knockdown or in ARCL2 cells resulted in distended Golgi cisternae, accumulation of abnormal lysosomes and multivesicular bodies. Immunostaining of ARCL2 cells showed the accumulation of tropoelastin (TE) in the Golgi and in large, abnormal intracellular and extracellular aggregates. Pulse-chase studies confirmed impaired secretion and increased intracellular retention of TE, and insoluble elastin assays showed significantly reduced extracellular deposition of mature elastin. Fibrillin-1 microfibril assembly and secreted lysyl oxidase activity were normal in ARCL2 cells. TUNEL staining demonstrated increased rates of apoptosis in ARCL2 cell cultures. We conclude that loss-of-function mutations in ATP6V0A2 lead to TE aggregation in the Golgi, impaired clearance of TE aggregates and increased apoptosis of elastogenic cells.
Das, S., Levinson, B., Vulpe, C., Whitney, S., Gitschier, J. and Packman, S. (1995) Similar splicing mutations of the Menkes/mottled copper-transporting ATPase gene in occipital horn syndrome and the blotchy mouse. Am. J. Hum. Genet., 56, 570-576.
Graul-Neumann, L.M., Hausser, I., Essayie, M., Rauch, A. and Kraus, C. (2008) Highly variable cutis laxa resulting from a dominant splicing mutation of the elastin gene. Am. J. Med. Genet. A, 146, 977-983.
Rodriguez-Revenga, L., Iranzo, P., Badenas, C., Puig, S., Carrio, A. and Mila, M. (2004) A novel elastin gene mutation resulting in an autosomal dominant form of cutis laxa. Arch. Dermatol., 140, 1135-1139.
Szabo, Z., Crepeau, M.W., Mitchell, A.L., Stephan, M.J., Puntel, R.A., Loke, K.Y., Kirk, R.C. and Urban, Z. (2006) Aortic aneurysmal disease and cutis laxa caused by defects in the elastin gene. J. Med. Genet. 43, 255-258.
Tassabehji, M., Metcalfe, K., Hurst, J., Ashcroft, G.S., Kielty, C., Wilmot, C., Donnai, D., Read, A.P. and Jones, C.J. (1998) An elastin gene mutation producing abnormal tropoelastin and abnormal elastic fibres in a patient with autosomal dominant cutis laxa. Hum. Mol. Genet., 7, 1021-1028.
Urban, Z., Gao, J., Pope, F.M. and Davis, E.C. (2005) Autosomal dominant cutis laxa with severe lung disease: Synthesis and matrix deposition of mutant tropoelastin. J. Invest. Dermatol., 124, 1193-1199.
Zhang, M.C., He, L., Giro, M., Yong, S.L., Tiller, G.E. and Davidson, J.M. (1999) Cutis laxa arising from frameshift mutations in exon 30 of the elastin gene (ELN). J. Biol. Chem., 274, 981-986.
Claus, S., Fischer, J., Megarbane, H., Megarbane, A., Jobard, F., Debret, R., Peyrol, S., Saker, S., Devillers, M., Sommer, P. et al. (2008) A p.C217R mutation in fibulin-5 from cutis laxa patients is associated with incomplete extracellular matrix formation in a skin equivalent model. J. Invest. Dermatol., 128, 1442-1450.
Loeys, B., Van Maldergem, L., Mortier, G., Coucke, P., Gerniers, S., Naeyaert, J.M. and De Paepe, A. (2002) Homozygosity for a missense mutation in fibulin-5 (FBLN5) results in a severe form of cutis laxa. Hum. Mol. Genet., 11, 2113-2118.
Dasouki, M., Markova, D., Garola, R., Sasaki, T., Charbonneau, N.L., Sakai, L.Y. and Chu, M.L. (2007) Compound heterozygous mutations in fibulin-4 causing neonatal lethal pulmonary artery occlusion, aortic aneurysm, arachnodactyly, and mild cutis laxa. Am. J. Med. Genet. A 143, 2635-2641.
Hucthagowder, V., Sausgruber, N., Kim, K.H., Angle, B., Marmorstein, L.Y. and Urban, Z. (2006) Fibulin-4: A novel gene for an autosomal recessive cutis laxa syndrome. Am. J. Hum. Genet., 78, 1075-1080.
Morava, E., Lefeber, D.J., Urban, Z., de Meirleir, L., Meinecke, P., Gillessen Kaesbach, G., Sykut-Cegielska, J., Adamowicz, M., Salafsky, I., Ranells, J. et al. (2008) Defining the phenotype in an autosomal recessive cutis laxa syndrome with a combined congenital defect of glycosylation. Eur. J. Hum. Genet., 16, 28-35.
Freeze, H.H. (2006) Genetic defects in the human glycome. Nat. Rev. Genet., 7, 537-551.
Foulquier, F., Ungar, D., Reynders, E., Zeevaert, R., Mills, P., Garcia-Silva, M.T., Briones, P., Winchester, B., Morelle, W., Krieger, M. et al. (2007) A new inborn error of glycosylation due to a Cog8 deficiency reveals a critical role for the Cog1-Cog8 interaction in COG complex formation. Hum. Mol. Genet., 16, 717-730.
Foulquier, F., Vasile, E., Schollen, E., Callewaert, N., Raemaekers, T., Quelhas, D., Jaeken, J., Mills, P., Winchester, B., Krieger, M. et al. (2006) Conserved oligomeric Golgi complex subunit 1 deficiency reveals a previously uncharacterized congenital disorder of glycosylation type II. Proc. Natl Acad. Sci. USA, 103, 3764-3769.
Wu, X., Steet, R.A., Bohorov, O., Bakker, J., Newell, J., Krieger, M., Spaapen, L., Kornfeld, S. and Freeze, H.H. (2004) Mutation of the COG complex subunit gene COG7 causes a lethal congenital disorder. Nat. Med., 10, 518-523.
Morava, E., Wopereis, S., Coucke, P., Gillessen-Kaesbach, G., Voit, T., Smeitink, J., Wevers, R. and Grunewald, S. (2005) Defective protein glycosylation in patients with cutis laxa syndrome. Eur. J. Hum. Genet., 13, 414-421.
Kornak, U., Reynders, E., Dimopoulou, A., van Reeuwijk, J., Fischer, B., Rajab, A., Budde, B., Nurnberg, P., Foulquier, F., Lefeber, D. et al. (2008) Impaired glycosylation and cutis laxa caused by mutations in the vesicular H-1-ATPase subunit ATP6V0A2. Nat. Genet., 40 32-34.
Van Maldergem, L., Yuksel-Apak, M., Kayserili, H., Seemanova, E., Giurgea, S., Basel-Vanagaite, L., Leao-Teles, E., Vigneron, J., Foulon, M., Greally, M. et al. (2008) Cobblestone-like brain dysgenesis and altered glycosylation in congenital cutis laxa, Debre type. Neurology, 71, 1602-1608.
Peralvarez-Marin, A., Lorenz-Fonfria, V.A., Simon-Vazquez, R., Gomariz, M., Meseguer, I., Querol, E. and Padros, E. (2008) Influence of proline on the thermostability of the active site and membrane arrangement of transmembrane proteins. Biophys. J., 95, 4384-4395.
Milewicz, D.M., Urban, Z. and Boyd, C. (2000) Genetic disorders of the elastic fiber system. Matrix Biol., 19, 471-480.
Kielty, C.M. (2006) Elastic fibres in health and disease. Expert Rev. Mol. Med., 8, 1-23.
Kozel, B.A., Rongish, B.J., Czirok, A., Zach, J., Little, C.D., Davis, E.C., Knutsen, R.H., Wagenseil, J.E., Levy, M.A. and Mecham, R.P. (2006) Elastic fiber formation: A dynamic view of extracellular matrix assembly using timer reporters. J. Cell. Physiol., 207, 87-96.
Davis, E.C. and Mecham, R.P. (1998) Intracellular trafficking of tropoelastin. Matrix Biol., 17, 245-254.
Davis, E.C., Broekelmann, T.J., Ozawa, Y. and Mecham, R.P. (1998) Identification of tropoelastin as a ligand for the 65-kD FK506-binding protein, FKBP65, in the secretory pathway. J. Cell. Biol., 140 295-303.
Urban, Z., Riazi, S., Seidl, T.L., Katahira, J., Smoot, L.B., Chitayat, D., Boyd, C.D. and Hinek, A. (2002) Connection between elastin haploinsufficiency and increased cell proliferation in patients with supravalvular aortic stenosis and Williams-Beuren syndrome. Am. J. Hum. Genet., 71, 30-44.
Madsen, E.C. and Gitlin, J.D. (2008) Zebrafish mutants calamity and catastrophe define critical pathways of gene-nutrient interactions in developmental copper metabolism. PLoS Genet., 4, e1000261.
Dettmer, J., Hong-Hermesdorf, A., Stierhof, Y.D. and Schumacher, K. (2006) Vacuolar H+-ATPase activity is required for endocytic and secretory trafficking in Arabidopsis. Plant Cell, 18, 715-730.
Taupenot, L., Harper, K.L. and O'Connor, D.T. (2005) Role of H-1-ATPase-mediated acidification in sorting and release of the regulated secretory protein chromogranin A: Evidence for a vesiculogenic function. J. Biol. Chem., 280, 3885-3897.
Tawfeek, H.A. and Abou-Samra, A.B. (2004) Important role for the V-type H(+)-ATPase and the Golgi apparatus in the recycling of PTH/PTHrP receptor. Am. J. Physiol. Endocrinol. Metab., 286, E704-E710.
Kinsey, R., Williamson, M.R., Chaudhry, S., Mellody, K.T., McGovern, A., Takahashi, S., Shuttleworth, C.A. and Kielty, C.M. (2008) Fibrillin-1 microfibril deposition is dependent on fibronectin assembly. J. Cell. Sci., 121, 2696-2704.
Sabatier, L., Chen, D., Fagotto-Kaufmann, C., Hubmacher, D., McKee, M.D., Annis, D.S., Mosher, D.F. and Reinhardt, D.P. (2009) Fibrillin assembly requires fibronectin. Mol. Biol. Cell, 20, 846-858.
Gansner, J.M., Mendelsohn, B.A., Hultman, K.A., Johnson, S.L. and Gitlin, J.D. (2007) Essential role of lysyl oxidases in notochord development. Dev. Biol., 307, 202-213.
Vrhovski, B., Jensen, S. and Weiss, A.S. (1997) Coacervation characteristics of recombinant human tropoelastin. Eur. J. Biochem. 250, 92-98.
Zeuzem, S., Feick, P., Zimmermann, P., Haase, W., Kahn, R.A. and Schulz, I. (1992) Intravesicular acidification correlates with binding of ADP-ribosylation factor to microsomal membranes. Proc. Natl Acad. Sci. USA, 89, 6619-6623.
Hinek, A., Pshezhetsky, A.V., von Itzstein, M. and Starcher, B. (2006) Lysosomal sialidase (neuraminidase-1) is targeted to the cell surface in a multiprotein complex that facilitates elastic fiber assembly. J. Biol. Chem., 281, 3698-3710.
Starcher, B., d'Azzo, A., Keller, P.W., Rao, G.K., Nadarajah, D. and Hinek, A. (2008) Neuraminidase-1 is required for the normal assembly of elastic fibers. Am. J. Physiol. Lung Cell. Mol. Physiol., 295 L637-L647.
Herrmann, B.G. and Frischauf, A.M. (1987) Isolation of genomic DNA. Methods Enzymol., 152, 180-183.
Wopereis, S., Morava, E., Grunewald, S., Adamowicz, M., Huijben, K.M., Lefeber, D.J. and Wevers, R.A. (2005) Patients with unsolved congenital disorders of glycosylation type II can be subdivided in six distinct biochemical groups. Glycobiology, 15, 1312-1319.
Davis, E.C. (1993) Smooth muscle cell to elastic lamina connections in developing mouse aorta. Role in aortic medial organization. Lab. Invest., 68, 89-99.
Palamakumbura, A.H. and Trackman, P.C. (2002) A fluorometric assay for detection of lysyl oxidase enzyme activity in biological samples. Anal. Biochem., 300, 245-251.
Hinek, A. and Rabinovitch, M. (1993) The ductus arteriosus migratory smooth muscle cell phenotype processes tropoelastin to a 52-kDa product associated with impaired assembly of elastic laminae. J. Biol. Chem. 268, 1405-1413.
