A single-cysteine mutant and chimeras of essential Leishmania Erv can complement the loss of Erv1 but not of Mia40 in yeast
| Autor: | Kai Hell, Linda Liedgens, Alexandra Stiegler, Margarida Duarte, Maike Eberhardt, Marcel Deponte, Ulrike Wirth, Sandra Specht, Ana M. Tomás |
|---|---|
| Přispěvatelé: | Instituto de Investigação e Inovação em Saúde |
| Jazyk: | angličtina |
| Rok vydání: | 2018 |
| Předmět: |
0301 basic medicine
Intermembrane space Protein Folding GSH reduced glutathione Mitochondrial intermembrane space Clinical Biochemistry Mutant Protozoan Proteins Protozoan Proteins / metabolismo medicine.disease_cause Mitochondrial Membrane Transport Proteins Biochemistry Lt Leishmania tarentolae Saccharomyces cerevisiae Proteins / genetics Mitochondrial Precursor Protein Import Complex Proteins Oxidoreductases Acting on Sulfur Group Donors Mitochondrial Proteins /genetics Leishmaniasis lcsh:QH301-705.5 Oxidative protein folding Leishmania Mutation lcsh:R5-920 Leishmaniasis / metabolismo Leishmania / genetics Mia40 Mitochondrial Membrane Transport Proteins / genetics Cell biology Mitochondria Complementation Saccharomyces cerevisiae / genetics lcsh:Medicine (General) Leishmania / pathogenicity Research Paper Sc Saccharomyces cerevisiae CHCHD4 Saccharomyces cerevisiae Proteins Erv Protein domain Saccharomyces cerevisiae Biology Leishmaniasis / parasitology Mitochondrial Proteins 03 medical and health sciences Leishmaniasis / genetics Protein Domains medicine Animals Humans IMS mitochondrial intermembrane space Pf Plasmodium falciparum Cysteine Protozoan Proteins / genetics Organic Chemistry Li Leishmania infantum Mitochondria / metabolismo biology.organism_classification Yeast Mitochondria / genetics Disease Models Animal 030104 developmental biology lcsh:Biology (General) Cysteine / genetics Oxidoreductases Acting on Sulfur Group Donors / genetics ALR |
| Zdroj: | Redox Biology, Vol 15, Iss C, Pp 363-374 (2018) Repositório Científico de Acesso Aberto de Portugal Repositório Científico de Acesso Aberto de Portugal (RCAAP) instacron:RCAAP Redox Biology |
| ISSN: | 2213-2317 |
| Popis: | Mia40/CHCHD4 and Erv1/ALR are essential for oxidative protein folding in the mitochondrial intermembrane space of yeast and mammals. In contrast, many protists, including important apicomplexan and kinetoplastid parasites, lack Mia40. Furthermore, the Erv homolog of the model parasite Leishmania tarentolae (LtErv) was shown to be incompatible with Saccharomyces cerevisiae Mia40 (ScMia40). Here we addressed structure-function relationships of ScErv1 and LtErv as well as their compatibility with the oxidative protein folding system in yeast using chimeric, truncated, and mutant Erv constructs. Chimeras between the N-terminal arm of ScErv1 and a variety of truncated LtErv constructs were able to rescue yeast cells that lack ScErv1. Yeast cells were also viable when only a single cysteine residue was replaced in LtErvC17S. Thus, the presence and position of the C-terminal arm and the kinetoplastida-specific second (KISS) domain of LtErv did not interfere with its functionality in the yeast system, whereas a relatively conserved cysteine residue before the flavodomain rendered LtErv incompatible with ScMia40. The question whether parasite Erv homologs might also exert the function of Mia40 was addressed in another set of complementation assays. However, neither the KISS domain nor other truncated or mutant LtErv constructs were able to rescue yeast cells that lack ScMia40. The general relevance of Erv and its candidate substrate small Tim1 was analyzed for the related parasite L. infantum. Repeated unsuccessful knockout attempts suggest that both genes are essential in this human pathogen and underline the potential of mitochondrial protein import pathways for future intervention strategies. Graphical abstract fx1 |
| Databáze: | OpenAIRE |
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