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Yeast model analysis of novel polymerase gamma variants found in patients with autosomal recessive mitochondrial disease

Replication of the mitochondrial genome depends on the single DNA polymerase (pol gamma). Mutations in the POLG gene, encoding the catalytic subunit of the human polymerase gamma, have been linked to a wide variety of mitochondrial disorders that show remarkable heterogeneity, with more than 200 seq...

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Autores principales: Kaliszewska, Magdalena, Kruszewski, Jakub, Kierdaszuk, Biruta, Kostera-Pruszczyk, Anna, Nojszewska, Monika, Łusakowska, Anna, Vizueta, Joel, Sabat, Dorota, Lutyk, Dorota, Lower, Michał, Piekutowska-Abramczuk, Dorota, Kaniak-Golik, Aneta, Pronicka, Ewa, Kamińska, Anna, Bartnik, Ewa, Golik, Paweł, Tońska, Katarzyna
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Springer Berlin Heidelberg 2015
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4529462/
https://www.ncbi.nlm.nih.gov/pubmed/26077851
http://dx.doi.org/10.1007/s00439-015-1578-x
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author Kaliszewska, Magdalena
Kruszewski, Jakub
Kierdaszuk, Biruta
Kostera-Pruszczyk, Anna
Nojszewska, Monika
Łusakowska, Anna
Vizueta, Joel
Sabat, Dorota
Lutyk, Dorota
Lower, Michał
Piekutowska-Abramczuk, Dorota
Kaniak-Golik, Aneta
Pronicka, Ewa
Kamińska, Anna
Bartnik, Ewa
Golik, Paweł
Tońska, Katarzyna
author_facet Kaliszewska, Magdalena
Kruszewski, Jakub
Kierdaszuk, Biruta
Kostera-Pruszczyk, Anna
Nojszewska, Monika
Łusakowska, Anna
Vizueta, Joel
Sabat, Dorota
Lutyk, Dorota
Lower, Michał
Piekutowska-Abramczuk, Dorota
Kaniak-Golik, Aneta
Pronicka, Ewa
Kamińska, Anna
Bartnik, Ewa
Golik, Paweł
Tońska, Katarzyna
author_sort Kaliszewska, Magdalena
collection PubMed
description Replication of the mitochondrial genome depends on the single DNA polymerase (pol gamma). Mutations in the POLG gene, encoding the catalytic subunit of the human polymerase gamma, have been linked to a wide variety of mitochondrial disorders that show remarkable heterogeneity, with more than 200 sequence variants, often very rare, found in patients. The pathogenicity and dominance status of many such mutations remain, however, unclear. Remarkable structural and functional conservation of human POLG and its S. cerevisiae ortholog (Mip1p) led to the development of many successful yeast models, enabling to study the phenotype of putative pathogenic mutations. In a group of patients with suspicion of mitochondrial pathology, we identified five novel POLG sequence variants, four of which (p.Arg869Ter, p.Gln968Glu, p.Thr1053Argfs*6, and p.Val1106Ala), together with one previously known but uncharacterised variant (p.Arg309Cys), were amenable to modelling in yeast. Familial analysis indicated causal relationship of these variants with disease, consistent with autosomal recessive inheritance. To investigate the effect of these sequence changes on mtDNA replication, we obtained the corresponding yeast mip1 alleles (Arg265Cys, Arg672Ter, Arg770Glu, Thr809Ter, and Val863Ala, respectively) and tested their effect on mitochondrial genome stability and replication fidelity. For three of them (Arg265Cys, Arg672Ter, and Thr809Ter), we observed a strong, partially dominant phenotype of a complete loss of functional mtDNA, whereas the remaining two led to partial mtDNA depletion and significant increase in point mutation frequencies. These results show good correlation with the severity of symptoms observed in patients and allow to establish these variants as pathogenic mutations. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (doi:10.1007/s00439-015-1578-x) contains supplementary material, which is available to authorized users.
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spelling pubmed-45294622015-08-11 Yeast model analysis of novel polymerase gamma variants found in patients with autosomal recessive mitochondrial disease Kaliszewska, Magdalena Kruszewski, Jakub Kierdaszuk, Biruta Kostera-Pruszczyk, Anna Nojszewska, Monika Łusakowska, Anna Vizueta, Joel Sabat, Dorota Lutyk, Dorota Lower, Michał Piekutowska-Abramczuk, Dorota Kaniak-Golik, Aneta Pronicka, Ewa Kamińska, Anna Bartnik, Ewa Golik, Paweł Tońska, Katarzyna Hum Genet Original Investigation Replication of the mitochondrial genome depends on the single DNA polymerase (pol gamma). Mutations in the POLG gene, encoding the catalytic subunit of the human polymerase gamma, have been linked to a wide variety of mitochondrial disorders that show remarkable heterogeneity, with more than 200 sequence variants, often very rare, found in patients. The pathogenicity and dominance status of many such mutations remain, however, unclear. Remarkable structural and functional conservation of human POLG and its S. cerevisiae ortholog (Mip1p) led to the development of many successful yeast models, enabling to study the phenotype of putative pathogenic mutations. In a group of patients with suspicion of mitochondrial pathology, we identified five novel POLG sequence variants, four of which (p.Arg869Ter, p.Gln968Glu, p.Thr1053Argfs*6, and p.Val1106Ala), together with one previously known but uncharacterised variant (p.Arg309Cys), were amenable to modelling in yeast. Familial analysis indicated causal relationship of these variants with disease, consistent with autosomal recessive inheritance. To investigate the effect of these sequence changes on mtDNA replication, we obtained the corresponding yeast mip1 alleles (Arg265Cys, Arg672Ter, Arg770Glu, Thr809Ter, and Val863Ala, respectively) and tested their effect on mitochondrial genome stability and replication fidelity. For three of them (Arg265Cys, Arg672Ter, and Thr809Ter), we observed a strong, partially dominant phenotype of a complete loss of functional mtDNA, whereas the remaining two led to partial mtDNA depletion and significant increase in point mutation frequencies. These results show good correlation with the severity of symptoms observed in patients and allow to establish these variants as pathogenic mutations. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (doi:10.1007/s00439-015-1578-x) contains supplementary material, which is available to authorized users. Springer Berlin Heidelberg 2015-06-16 2015 /pmc/articles/PMC4529462/ /pubmed/26077851 http://dx.doi.org/10.1007/s00439-015-1578-x Text en © The Author(s) 2015 Open AccessThis article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made.
spellingShingle Original Investigation
Kaliszewska, Magdalena
Kruszewski, Jakub
Kierdaszuk, Biruta
Kostera-Pruszczyk, Anna
Nojszewska, Monika
Łusakowska, Anna
Vizueta, Joel
Sabat, Dorota
Lutyk, Dorota
Lower, Michał
Piekutowska-Abramczuk, Dorota
Kaniak-Golik, Aneta
Pronicka, Ewa
Kamińska, Anna
Bartnik, Ewa
Golik, Paweł
Tońska, Katarzyna
Yeast model analysis of novel polymerase gamma variants found in patients with autosomal recessive mitochondrial disease
title Yeast model analysis of novel polymerase gamma variants found in patients with autosomal recessive mitochondrial disease
title_full Yeast model analysis of novel polymerase gamma variants found in patients with autosomal recessive mitochondrial disease
title_fullStr Yeast model analysis of novel polymerase gamma variants found in patients with autosomal recessive mitochondrial disease
title_full_unstemmed Yeast model analysis of novel polymerase gamma variants found in patients with autosomal recessive mitochondrial disease
title_short Yeast model analysis of novel polymerase gamma variants found in patients with autosomal recessive mitochondrial disease
title_sort yeast model analysis of novel polymerase gamma variants found in patients with autosomal recessive mitochondrial disease
topic Original Investigation
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4529462/
https://www.ncbi.nlm.nih.gov/pubmed/26077851
http://dx.doi.org/10.1007/s00439-015-1578-x
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