Loss-of-function mutations in ATP6V0A2 impair vesicular trafficking, tropoelastin secretion and cell survival

Vishwanathan Hucthagowder, Eva Morava, Uwe Kornak, Dirk J. Lefeber, Björn Fischer, Aikaterini Dimopoulou, Annika Aldinger, Jiwon Choi, Elaine C. Davis, Dianne N. Abuelo, Maciej Adamowicz, Jumana Al-Aama, Lina Basel-Vanagaite, Bridget Fernandez, Marie T. Greally, Gabriele Gillessen-Kaesbach, Hulya Kayserili, Emmanuelle Lemyre, Mustafa Tekin, Seval TürkmenBeyhan Tuysuz, Berrin Yüksel-Konuk, Stefan Mundlos, Lionel Van Maldergem, Ron A. Wevers, Zsolt Urban

Research output: Contribution to journalArticle

73 Citations (Scopus)

Abstract

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.

Original languageEnglish
Pages (from-to)2149-2165
Number of pages17
JournalHuman Molecular Genetics
Volume18
Issue number12
DOIs
StatePublished - Jun 4 2009
Externally publishedYes

Fingerprint

Tropoelastin
Cell Survival
Mutation
Elastin
Nonsense Mediated mRNA Decay
Protein-Lysine 6-Oxidase
Apoptosis
Multivesicular Bodies
Microfibrils
Proton Pumps
Nonsense Codon
In Situ Nick-End Labeling
Mucins
Missense Mutation
Lysosomes
DNA Sequence Analysis
Glycosylation
Connective Tissue
Small Interfering RNA
Adenosine Triphosphatases

ASJC Scopus subject areas

  • Genetics
  • Genetics(clinical)
  • Molecular Biology

Cite this

Hucthagowder, V., Morava, E., Kornak, U., Lefeber, D. J., Fischer, B., Dimopoulou, A., ... Urban, Z. (2009). Loss-of-function mutations in ATP6V0A2 impair vesicular trafficking, tropoelastin secretion and cell survival. Human Molecular Genetics, 18(12), 2149-2165. https://doi.org/10.1093/hmg/ddp148

Loss-of-function mutations in ATP6V0A2 impair vesicular trafficking, tropoelastin secretion and cell survival. / Hucthagowder, Vishwanathan; Morava, Eva; Kornak, Uwe; Lefeber, Dirk J.; Fischer, Björn; Dimopoulou, Aikaterini; Aldinger, Annika; Choi, Jiwon; Davis, Elaine C.; Abuelo, Dianne N.; Adamowicz, Maciej; Al-Aama, Jumana; Basel-Vanagaite, Lina; Fernandez, Bridget; Greally, Marie T.; Gillessen-Kaesbach, Gabriele; Kayserili, Hulya; Lemyre, Emmanuelle; Tekin, Mustafa; Türkmen, Seval; Tuysuz, Beyhan; Yüksel-Konuk, Berrin; Mundlos, Stefan; Van Maldergem, Lionel; Wevers, Ron A.; Urban, Zsolt.

In: Human Molecular Genetics, Vol. 18, No. 12, 04.06.2009, p. 2149-2165.

Research output: Contribution to journalArticle

Hucthagowder, V, Morava, E, Kornak, U, Lefeber, DJ, Fischer, B, Dimopoulou, A, Aldinger, A, Choi, J, Davis, EC, Abuelo, DN, Adamowicz, M, Al-Aama, J, Basel-Vanagaite, L, Fernandez, B, Greally, MT, Gillessen-Kaesbach, G, Kayserili, H, Lemyre, E, Tekin, M, Türkmen, S, Tuysuz, B, Yüksel-Konuk, B, Mundlos, S, Van Maldergem, L, Wevers, RA & Urban, Z 2009, 'Loss-of-function mutations in ATP6V0A2 impair vesicular trafficking, tropoelastin secretion and cell survival', Human Molecular Genetics, vol. 18, no. 12, pp. 2149-2165. https://doi.org/10.1093/hmg/ddp148
Hucthagowder V, Morava E, Kornak U, Lefeber DJ, Fischer B, Dimopoulou A et al. Loss-of-function mutations in ATP6V0A2 impair vesicular trafficking, tropoelastin secretion and cell survival. Human Molecular Genetics. 2009 Jun 4;18(12):2149-2165. https://doi.org/10.1093/hmg/ddp148
Hucthagowder, Vishwanathan ; Morava, Eva ; Kornak, Uwe ; Lefeber, Dirk J. ; Fischer, Björn ; Dimopoulou, Aikaterini ; Aldinger, Annika ; Choi, Jiwon ; Davis, Elaine C. ; Abuelo, Dianne N. ; Adamowicz, Maciej ; Al-Aama, Jumana ; Basel-Vanagaite, Lina ; Fernandez, Bridget ; Greally, Marie T. ; Gillessen-Kaesbach, Gabriele ; Kayserili, Hulya ; Lemyre, Emmanuelle ; Tekin, Mustafa ; Türkmen, Seval ; Tuysuz, Beyhan ; Yüksel-Konuk, Berrin ; Mundlos, Stefan ; Van Maldergem, Lionel ; Wevers, Ron A. ; Urban, Zsolt. / Loss-of-function mutations in ATP6V0A2 impair vesicular trafficking, tropoelastin secretion and cell survival. In: Human Molecular Genetics. 2009 ; Vol. 18, No. 12. pp. 2149-2165.
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abstract = "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.",
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AU - Lefeber, Dirk J.

AU - Fischer, Björn

AU - Dimopoulou, Aikaterini

AU - Aldinger, Annika

AU - Choi, Jiwon

AU - Davis, Elaine C.

AU - Abuelo, Dianne N.

AU - Adamowicz, Maciej

AU - Al-Aama, Jumana

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AU - Fernandez, Bridget

AU - Greally, Marie T.

AU - Gillessen-Kaesbach, Gabriele

AU - Kayserili, Hulya

AU - Lemyre, Emmanuelle

AU - Tekin, Mustafa

AU - Türkmen, Seval

AU - Tuysuz, Beyhan

AU - Yüksel-Konuk, Berrin

AU - Mundlos, Stefan

AU - Van Maldergem, Lionel

AU - Wevers, Ron A.

AU - Urban, Zsolt

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N2 - 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.

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