Biallelic Mutations in MRPS34 Lead to Instability of the Small Mitoribosomal Subunit and Leigh Syndrome

Nicole J. Lake, Bryn D. Webb, David A. Stroud, Tara R. Richman, Benedetta Ruzzenente, Alison G. Compton, Hayley S. Mountford, Juliette Pulman, Coralie Zangarelli, Marlene Rio, Nathalie Bodaert, Zahra Assouline, Mingma D. Sherpa, Eric E. Schadt, Sander M. Houten, James Byrnes, Elizabeth M. McCormick, Zarazuela Zolkipli-Cunningham, Katrina Haude, Zhancheng Zhang & 11 others Kyle Retterer, Renkui Bai, Sarah E. Calvo, Vamsi K. Mootha, John Christodoulou, Agnes Rötig, Aleksandra Filipovska, Ingrid Cristian, Marni J. Falk, Metodi D. Metodiev, David R. Thorburn

Research output: Contribution to journalArticleResearchpeer-review

18 Citations (Scopus)

Abstract

The synthesis of all 13 mitochondrial DNA (mtDNA)-encoded protein subunits of the human oxidative phosphorylation (OXPHOS) system is carried out by mitochondrial ribosomes (mitoribosomes). Defects in the stability of mitoribosomal proteins or mitoribosome assembly impair mitochondrial protein translation, causing combined OXPHOS enzyme deficiency and clinical disease. Here we report four autosomal-recessive pathogenic mutations in the gene encoding the small mitoribosomal subunit protein, MRPS34, in six subjects from four unrelated families with Leigh syndrome and combined OXPHOS defects. Whole-exome sequencing was used to independently identify all variants. Two splice-site mutations were identified, including homozygous c.321+1G>T in a subject of Italian ancestry and homozygous c.322−10G>A in affected sibling pairs from two unrelated families of Puerto Rican descent. In addition, compound heterozygous MRPS34 mutations were identified in a proband of French ancestry; a missense (c.37G>A [p.Glu13Lys]) and a nonsense (c.94C>T [p.Gln32]) variant. We demonstrated that these mutations reduce MRPS34 protein levels and the synthesis of OXPHOS subunits encoded by mtDNA. Examination of the mitoribosome profile and quantitative proteomics showed that the mitochondrial translation defect was caused by destabilization of the small mitoribosomal subunit and impaired monosome assembly. Lentiviral-mediated expression of wild-type MRPS34 rescued the defect in mitochondrial translation observed in skin fibroblasts from affected subjects, confirming the pathogenicity of MRPS34 mutations. Our data establish that MRPS34 is required for normal function of the mitoribosome in humans and furthermore demonstrate the power of quantitative proteomic analysis to identify signatures of defects in specific cellular pathways in fibroblasts from subjects with inherited disease.

Original languageEnglish
Pages (from-to)239-254
Number of pages16
JournalAmerican Journal of Human Genetics
Volume101
Issue number2
DOIs
Publication statusPublished - 3 Aug 2017

Keywords

  • Leigh syndrome
  • mitochondrial diseases
  • mitochondrial ribosome
  • mitochondrial translation
  • MRPS34
  • quantitative proteomics
  • respiratory chain
  • ribosome profiling
  • whole-exome sequencing

Cite this

Lake, N. J., Webb, B. D., Stroud, D. A., Richman, T. R., Ruzzenente, B., Compton, A. G., ... Thorburn, D. R. (2017). Biallelic Mutations in MRPS34 Lead to Instability of the Small Mitoribosomal Subunit and Leigh Syndrome. American Journal of Human Genetics, 101(2), 239-254. https://doi.org/10.1016/j.ajhg.2017.07.005
Lake, Nicole J. ; Webb, Bryn D. ; Stroud, David A. ; Richman, Tara R. ; Ruzzenente, Benedetta ; Compton, Alison G. ; Mountford, Hayley S. ; Pulman, Juliette ; Zangarelli, Coralie ; Rio, Marlene ; Bodaert, Nathalie ; Assouline, Zahra ; Sherpa, Mingma D. ; Schadt, Eric E. ; Houten, Sander M. ; Byrnes, James ; McCormick, Elizabeth M. ; Zolkipli-Cunningham, Zarazuela ; Haude, Katrina ; Zhang, Zhancheng ; Retterer, Kyle ; Bai, Renkui ; Calvo, Sarah E. ; Mootha, Vamsi K. ; Christodoulou, John ; Rötig, Agnes ; Filipovska, Aleksandra ; Cristian, Ingrid ; Falk, Marni J. ; Metodiev, Metodi D. ; Thorburn, David R. / Biallelic Mutations in MRPS34 Lead to Instability of the Small Mitoribosomal Subunit and Leigh Syndrome. In: American Journal of Human Genetics. 2017 ; Vol. 101, No. 2. pp. 239-254.
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title = "Biallelic Mutations in MRPS34 Lead to Instability of the Small Mitoribosomal Subunit and Leigh Syndrome",
abstract = "The synthesis of all 13 mitochondrial DNA (mtDNA)-encoded protein subunits of the human oxidative phosphorylation (OXPHOS) system is carried out by mitochondrial ribosomes (mitoribosomes). Defects in the stability of mitoribosomal proteins or mitoribosome assembly impair mitochondrial protein translation, causing combined OXPHOS enzyme deficiency and clinical disease. Here we report four autosomal-recessive pathogenic mutations in the gene encoding the small mitoribosomal subunit protein, MRPS34, in six subjects from four unrelated families with Leigh syndrome and combined OXPHOS defects. Whole-exome sequencing was used to independently identify all variants. Two splice-site mutations were identified, including homozygous c.321+1G>T in a subject of Italian ancestry and homozygous c.322−10G>A in affected sibling pairs from two unrelated families of Puerto Rican descent. In addition, compound heterozygous MRPS34 mutations were identified in a proband of French ancestry; a missense (c.37G>A [p.Glu13Lys]) and a nonsense (c.94C>T [p.Gln32∗]) variant. We demonstrated that these mutations reduce MRPS34 protein levels and the synthesis of OXPHOS subunits encoded by mtDNA. Examination of the mitoribosome profile and quantitative proteomics showed that the mitochondrial translation defect was caused by destabilization of the small mitoribosomal subunit and impaired monosome assembly. Lentiviral-mediated expression of wild-type MRPS34 rescued the defect in mitochondrial translation observed in skin fibroblasts from affected subjects, confirming the pathogenicity of MRPS34 mutations. Our data establish that MRPS34 is required for normal function of the mitoribosome in humans and furthermore demonstrate the power of quantitative proteomic analysis to identify signatures of defects in specific cellular pathways in fibroblasts from subjects with inherited disease.",
keywords = "Leigh syndrome, mitochondrial diseases, mitochondrial ribosome, mitochondrial translation, MRPS34, quantitative proteomics, respiratory chain, ribosome profiling, whole-exome sequencing",
author = "Lake, {Nicole J.} and Webb, {Bryn D.} and Stroud, {David A.} and Richman, {Tara R.} and Benedetta Ruzzenente and Compton, {Alison G.} and Mountford, {Hayley S.} and Juliette Pulman and Coralie Zangarelli and Marlene Rio and Nathalie Bodaert and Zahra Assouline and Sherpa, {Mingma D.} and Schadt, {Eric E.} and Houten, {Sander M.} and James Byrnes and McCormick, {Elizabeth M.} and Zarazuela Zolkipli-Cunningham and Katrina Haude and Zhancheng Zhang and Kyle Retterer and Renkui Bai and Calvo, {Sarah E.} and Mootha, {Vamsi K.} and John Christodoulou and Agnes R{\"o}tig and Aleksandra Filipovska and Ingrid Cristian and Falk, {Marni J.} and Metodiev, {Metodi D.} and Thorburn, {David R.}",
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Lake, NJ, Webb, BD, Stroud, DA, Richman, TR, Ruzzenente, B, Compton, AG, Mountford, HS, Pulman, J, Zangarelli, C, Rio, M, Bodaert, N, Assouline, Z, Sherpa, MD, Schadt, EE, Houten, SM, Byrnes, J, McCormick, EM, Zolkipli-Cunningham, Z, Haude, K, Zhang, Z, Retterer, K, Bai, R, Calvo, SE, Mootha, VK, Christodoulou, J, Rötig, A, Filipovska, A, Cristian, I, Falk, MJ, Metodiev, MD & Thorburn, DR 2017, 'Biallelic Mutations in MRPS34 Lead to Instability of the Small Mitoribosomal Subunit and Leigh Syndrome', American Journal of Human Genetics, vol. 101, no. 2, pp. 239-254. https://doi.org/10.1016/j.ajhg.2017.07.005

Biallelic Mutations in MRPS34 Lead to Instability of the Small Mitoribosomal Subunit and Leigh Syndrome. / Lake, Nicole J.; Webb, Bryn D.; Stroud, David A.; Richman, Tara R.; Ruzzenente, Benedetta; Compton, Alison G.; Mountford, Hayley S.; Pulman, Juliette; Zangarelli, Coralie; Rio, Marlene; Bodaert, Nathalie; Assouline, Zahra; Sherpa, Mingma D.; Schadt, Eric E.; Houten, Sander M.; Byrnes, James; McCormick, Elizabeth M.; Zolkipli-Cunningham, Zarazuela; Haude, Katrina; Zhang, Zhancheng; Retterer, Kyle; Bai, Renkui; Calvo, Sarah E.; Mootha, Vamsi K.; Christodoulou, John; Rötig, Agnes; Filipovska, Aleksandra; Cristian, Ingrid; Falk, Marni J.; Metodiev, Metodi D.; Thorburn, David R.

In: American Journal of Human Genetics, Vol. 101, No. 2, 03.08.2017, p. 239-254.

Research output: Contribution to journalArticleResearchpeer-review

TY - JOUR

T1 - Biallelic Mutations in MRPS34 Lead to Instability of the Small Mitoribosomal Subunit and Leigh Syndrome

AU - Lake, Nicole J.

AU - Webb, Bryn D.

AU - Stroud, David A.

AU - Richman, Tara R.

AU - Ruzzenente, Benedetta

AU - Compton, Alison G.

AU - Mountford, Hayley S.

AU - Pulman, Juliette

AU - Zangarelli, Coralie

AU - Rio, Marlene

AU - Bodaert, Nathalie

AU - Assouline, Zahra

AU - Sherpa, Mingma D.

AU - Schadt, Eric E.

AU - Houten, Sander M.

AU - Byrnes, James

AU - McCormick, Elizabeth M.

AU - Zolkipli-Cunningham, Zarazuela

AU - Haude, Katrina

AU - Zhang, Zhancheng

AU - Retterer, Kyle

AU - Bai, Renkui

AU - Calvo, Sarah E.

AU - Mootha, Vamsi K.

AU - Christodoulou, John

AU - Rötig, Agnes

AU - Filipovska, Aleksandra

AU - Cristian, Ingrid

AU - Falk, Marni J.

AU - Metodiev, Metodi D.

AU - Thorburn, David R.

PY - 2017/8/3

Y1 - 2017/8/3

N2 - The synthesis of all 13 mitochondrial DNA (mtDNA)-encoded protein subunits of the human oxidative phosphorylation (OXPHOS) system is carried out by mitochondrial ribosomes (mitoribosomes). Defects in the stability of mitoribosomal proteins or mitoribosome assembly impair mitochondrial protein translation, causing combined OXPHOS enzyme deficiency and clinical disease. Here we report four autosomal-recessive pathogenic mutations in the gene encoding the small mitoribosomal subunit protein, MRPS34, in six subjects from four unrelated families with Leigh syndrome and combined OXPHOS defects. Whole-exome sequencing was used to independently identify all variants. Two splice-site mutations were identified, including homozygous c.321+1G>T in a subject of Italian ancestry and homozygous c.322−10G>A in affected sibling pairs from two unrelated families of Puerto Rican descent. In addition, compound heterozygous MRPS34 mutations were identified in a proband of French ancestry; a missense (c.37G>A [p.Glu13Lys]) and a nonsense (c.94C>T [p.Gln32∗]) variant. We demonstrated that these mutations reduce MRPS34 protein levels and the synthesis of OXPHOS subunits encoded by mtDNA. Examination of the mitoribosome profile and quantitative proteomics showed that the mitochondrial translation defect was caused by destabilization of the small mitoribosomal subunit and impaired monosome assembly. Lentiviral-mediated expression of wild-type MRPS34 rescued the defect in mitochondrial translation observed in skin fibroblasts from affected subjects, confirming the pathogenicity of MRPS34 mutations. Our data establish that MRPS34 is required for normal function of the mitoribosome in humans and furthermore demonstrate the power of quantitative proteomic analysis to identify signatures of defects in specific cellular pathways in fibroblasts from subjects with inherited disease.

AB - The synthesis of all 13 mitochondrial DNA (mtDNA)-encoded protein subunits of the human oxidative phosphorylation (OXPHOS) system is carried out by mitochondrial ribosomes (mitoribosomes). Defects in the stability of mitoribosomal proteins or mitoribosome assembly impair mitochondrial protein translation, causing combined OXPHOS enzyme deficiency and clinical disease. Here we report four autosomal-recessive pathogenic mutations in the gene encoding the small mitoribosomal subunit protein, MRPS34, in six subjects from four unrelated families with Leigh syndrome and combined OXPHOS defects. Whole-exome sequencing was used to independently identify all variants. Two splice-site mutations were identified, including homozygous c.321+1G>T in a subject of Italian ancestry and homozygous c.322−10G>A in affected sibling pairs from two unrelated families of Puerto Rican descent. In addition, compound heterozygous MRPS34 mutations were identified in a proband of French ancestry; a missense (c.37G>A [p.Glu13Lys]) and a nonsense (c.94C>T [p.Gln32∗]) variant. We demonstrated that these mutations reduce MRPS34 protein levels and the synthesis of OXPHOS subunits encoded by mtDNA. Examination of the mitoribosome profile and quantitative proteomics showed that the mitochondrial translation defect was caused by destabilization of the small mitoribosomal subunit and impaired monosome assembly. Lentiviral-mediated expression of wild-type MRPS34 rescued the defect in mitochondrial translation observed in skin fibroblasts from affected subjects, confirming the pathogenicity of MRPS34 mutations. Our data establish that MRPS34 is required for normal function of the mitoribosome in humans and furthermore demonstrate the power of quantitative proteomic analysis to identify signatures of defects in specific cellular pathways in fibroblasts from subjects with inherited disease.

KW - Leigh syndrome

KW - mitochondrial diseases

KW - mitochondrial ribosome

KW - mitochondrial translation

KW - MRPS34

KW - quantitative proteomics

KW - respiratory chain

KW - ribosome profiling

KW - whole-exome sequencing

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U2 - 10.1016/j.ajhg.2017.07.005

DO - 10.1016/j.ajhg.2017.07.005

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JO - American Journal of Human Genetics

JF - American Journal of Human Genetics

SN - 0002-9297

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