Substitutions in conserved regions preceding and within the linker affect activity and flexibility of tRNase ZL, the long form of tRNase Z

Saoura, Makenzie; Pinnock, Kyla; Pujantell-Graell, Maria; Levinger, Louis

doi:10.1371/journal.pone.0186277

Cited by 4 publications

(9 citation statements)

References 18 publications

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“…In order to provide evidence for the pathogenicity of identified ELAC2 variants, we set out to study the RNase Z activity of the enzyme in the presence of the missense substitutions or the truncating variants resulting from frameshift mutations. Substitutions were introduced into the human ELAC2 cDNA (Genbank Acc# NM_018127.6) and the mutant proteins were expressed in baculovirus using insect SF9 cells [35]. Affinity-purified recombinant mutant proteins were tested using precursor mt-tRNA LeuUUR or mt-tRNA Ile as substrates.…”

Section: In Vitro Rnase Z Activity Of Mutant Elac2 Enzymesmentioning

confidence: 99%

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Mutations in ELAC2 associated with hypertrophic cardiomyopathy impair mitochondrial tRNA 3’-end processing

Saoura

Powell

Kopajtich

et al. 2018

Preprint

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Dysfunction of mitochondrial gene expression, caused by mutations in either the mitochondrial or nuclear genomes, is associated with a diverse group of human disorders characterized by impaired mitochondrial respiration. Within this group, an increasing number of mutations have been identified in nuclear genes involved in mitochondrial RNA metabolism. For instance, pathogenic mutations have been identified in the genes encoding enzymes involved in the precursor transcript processing, including ELAC2. The ELAC2 gene codes for the mitochondrial RNase Z, which is responsible for endonucleolytic cleavage of the 3' ends of mitochondrial pre-tRNAs. Here, we report the identification of sixteen novel ELAC2 variants in individuals presenting with mitochondrial respiratory chain deficiency, hypertrophic cardiomyopathy and lactic acidosis. We provided further evidence for the pathogenicity of the three previously reported variants by studying the RNase Z activity in an in vitro system and applied this recombinant system to investigate all novel missense variants, confirming the pathogenic role of these new ELAC2 mutations. We also modelled the residues affected by missense mutation in solved RNase Z structures, providing insight into enzyme structure and function. Finally, we show that primary fibroblasts from the individuals with novel ELAC2 variants have elevated levels of unprocessed mitochondrial RNA precursors. Our study thus broadly confirms the correlation of ELAC2 variants with severe infantile-onset forms of hypertrophic cardiomyopathy and mitochondrial respiratory chain dysfunction. One rare missense variant associated with the occurrence of prostate cancer (p.Arg781His) impairs the mitochondrial RNase Z activity of ELAC2, possibly indicating a functional link between tumorigenesis and mitochondrial RNA metabolism.

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Section: In Vitro Rnase Z Activity Of Mutant Elac2 Enzymesmentioning

confidence: 99%

“…ELAC2 mutations and gene/protein structure (A) Gene structure of ELAC2 with known protein domains (as defined in[35]) of the gene product and localization of amino acid residues and splice sites (variants indicated in blue) affected by mutations. Intronic regions are not drawn to scale.…”

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Mutations in ELAC2 associated with hypertrophic cardiomyopathy impair mitochondrial tRNA 3’-end processing

Saoura

Powell

Kopajtich

et al. 2018

Preprint

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“…C) Estrutura cristalográfica das metalo-β-lactamase contendo dois íons de cinco no centro catalítico (PDB: 1QH3 cadeia Adaptada de (Schilling et al, 2005)). curta (RNase Z S , entre 250 e 400 resíduos de aminoácidos) presente em bactérias, arqueias e alguns eucariotos e a variante longa (RNase Z L , entre 750 e 900) encontradas exclusivamente em eucariotos (Ma et al, 2017;Saoura et al, 2017). A variante curta, RNase Z S , age como homodímero e além dos motivos I-V, tem um braço flexível (exosite) entre os motivos III e IV que permite a interação com os tRNA (Figura 1.1A).…”

Section: Processamento De Rnasunclassified

“…A variante longa, RNase Z L , foi identificada como monômero em solução e propõe-se que surgiu como uma duplicação em tandem da variante curta ou mediante a fusão de dois genes distintos: a variante curta de RNase Z e outro gene com similaridade de sequência a variante curta (Fischer et al, 2012). O domínio C-terminal (CTD) das RNase Z L compartilha identidade de sequência com as RNase Z S , mas as sequências do domínio N-terminal (NTD) divergem da família proteica, mesmo assim ele é considerado como um dobramento β-lactamase (Ma et al, 2017;Saoura et al, 2017).…”

Section: Processamento De Rnasunclassified

“…Para estudar a conservação dos motivos das MBL no produto proteico do gene VNG_RS05855 realizamos o alinhamento múltiplo, incluindo sequências proteicas das RNases Z de organismos procariotos nas que se demonstrou atividade ribonucleolítica, tanto in vivo como in vitro, (Schierling et al, 2002;Vogel et al, 2005;Schilling et al, 2005) (SCHIERLING et al, 2002VOGEL;SPA;, o motivo carboxi-terminal da RNase Z L de Saccharomyces cerevisiae que conserva os resíduos catalíticos (Ma et al, 2017;Saoura et al, 2017) Apesar da proteína codificada pelo gene VNG_RS05855 ter poucas chances de reconhecer pré-tRNAs como alvos, não podemos descartar a sua possível atividade ribonucleolítica. Esta ideia é plausível ao analisar o alinhamento da sequência de aminoácidos do gene VNG_RS05855 com o motivo catalítico carboxi-terminal da RNase Z L de S. cerevisiae (Sce*) e observar que compartilha as características de conservação dos aminoácidos do domínio funcional e ausência do exosite (Saoura et al, 2017;Ma et al, 2017). Outra perspectiva para analisar a possível atividade ribonucleolítica do produto proteico do gene VNG_RS05855 são os relatos da capacidade da RNase Z de processar outros RNAs mRNA (Redko, Li de Lasierra-Gallay e Condon, 2007;Hölzle et al, 2008).…”

Section: A Sequência Proteica Do Gene Vng_rs05855 Conserva Os Motivosunclassified

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Regulação pós-transcricional em Halobacterium salinarum NRC-1: caracterização do gene VNG_RS05855

Marrero-Gutiérrez¹

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Agradeço à Prof a. Dr a. Tie Koide pela oportunidade de integrar seu grupo de pesquisa, pelo incentivo, pelo apoio e por estar sempre disposta a discussões. Agradeço pelos conselhos e por me mostrar o caminho de como fazer e escrever ciência. Obrigada pela paciência, orientação, confiança e contribuição na minha formação profissional! Agradeço ao Prof. Dr. Ricardo N. Z. Vêncio pela orientação, pelas constantes discussões e questionamentos, pelo apoio, pela confiança. Agradeço por me mostrar o útil de ser criterioso, focado, disciplinado e curioso na hora de fazer ciência, mas principalmente agradeço por ser amigo. Agradeço ao Prof. Dr. Fernando Barroso, pela oportunidade de vir para o Brasil. Agradeço a todo o corpo acadêmico e aos funcionários administrativos (antigos e atuais) do Programa de Interunidades de Pós-Graduação em Bioinformática, aos funcionários do

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Mutations inELAC2associated with hypertrophic cardiomyopathy impair mitochondrial tRNA 3′‐end processing

et al. 2019

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Mutations in either the mitochondrial or nuclear genomes are associated with a diverse group of human disorders characterized by impaired mitochondrial respiration. Within this group, an increasing number of mutations have been identified in nuclear genes involved in mitochondrial RNA metabolism, including ELAC2. The ELAC2 gene codes for the mitochondrial RNase Z, responsible for endonucleolytic cleavage of the 3′ ends of mitochondrial pre‐tRNAs. Here, we report the identification of 16 novel ELAC2 variants in individuals presenting with mitochondrial respiratory chain deficiency, hypertrophic cardiomyopathy (HCM), and lactic acidosis. We provide evidence for the pathogenicity of the novel missense variants by studying the RNase Z activity in an in vitro system. We also modeled the residues affected by a missense mutation in solved RNase Z structures, providing insight into enzyme structure and function. Finally, we show that primary fibroblasts from the affected individuals have elevated levels of unprocessed mitochondrial RNA precursors. Our study thus broadly confirms the correlation of ELAC2 variants with severe infantile‐onset forms of HCM and mitochondrial respiratory chain dysfunction. One rare missense variant associated with the occurrence of prostate cancer (p.Arg781His) impairs the mitochondrial RNase Z activity of ELAC2, suggesting a functional link between tumorigenesis and mitochondrial RNA metabolism

show abstract

Substitutions in conserved regions preceding and within the linker affect activity and flexibility of tRNase ZL, the long form of tRNase Z

Cited by 4 publications

References 18 publications

Mutations in ELAC2 associated with hypertrophic cardiomyopathy impair mitochondrial tRNA 3’-end processing

Mutations in ELAC2 associated with hypertrophic cardiomyopathy impair mitochondrial tRNA 3’-end processing

Regulação pós-transcricional em Halobacterium salinarum NRC-1: caracterização do gene VNG_RS05855

Mutations inELAC2associated with hypertrophic cardiomyopathy impair mitochondrial tRNA 3′‐end processing

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