biostudies-literatureTurra GLDeutscher Akademischer AustauschdienstDeutsche Forschungsgemeinschafte0080921https://www.ebi.ac.uk/biostudies/studies/S-EPMC8557897biostudies-literatureUnknown9(2)Import and oxidative folding of proteins in the mitochondrial intermembrane space differ among eukaryotic lineages. While opisthokonts such as yeast rely on the receptor and oxidoreductase Mia40 in combination with the Mia40:cytochrome <i>c</i> oxidoreductase Erv, kinetoplastid parasites and other Excavata/Discoba lack Mia40 but have a functional Erv homologue. Whether excavate Erv homologues rely on a Mia40 replacement or directly interact with imported protein substrates remains controversial. Here, we used the CRISPR-Cas9 system to generate a set of tagged and untagged homozygous mutants of <i>LTERV</i> from the kinetoplastid model parasite Leishmania tarentolae. Modifications of the shuttle cysteine motif of <i>Lt</i>Erv were lethal, whereas replacement of clamp residue Cys<sup>17</supMicrobiology spectrum<i>In Vivo</i> Structure-Function Analysis and Redox Interactomes of Leishmania tarentolae Erv.PMC8557897DE 1431/10-2DE 1431/10-1DE 1431/9-1Liedgens LSchroda MDeponte MTurra GLVilurbina Perez JKoncarevic SZimmer DMuhlhaus TSommer FSchneider Lfalse<i>In Vivo</i> Structure-Function Analysis and Redox Interactomes of Leishmania tarentolae Erv.Import and oxidative folding of proteins in the mitochondrial intermembrane space differ among eukaryotic lineages. While opisthokonts such as yeast rely on the receptor and oxidoreductase Mia40 in combination with the Mia40:cytochrome <i>c</i> oxidoreductase Erv, kinetoplastid parasites and other Excavata/Discoba lack Mia40 but have a functional Erv homologue. Whether excavate Erv homologues rely on a Mia40 replacement or directly interact with imported protein substrates remains controversial. Here, we used the CRISPR-Cas9 system to generate a set of tagged and untagged homozygous mutants of <i>LTERV</i> from the kinetoplastid model parasite Leishmania tarentolae. Modifications of the shuttle cysteine motif of <i>Lt</i>Erv were lethal, whereas replacement of clamp residue Cys<sup>17</sup2021-01-01T00:00:00Z2021 Oct2026-06-15T04:52:34.339Z2022-02-11T12:40:28.393ZS-EPMC85578973458598810.1128/Spectrum.00809-21