{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,10,12]],"date-time":"2025-10-12T03:08:33Z","timestamp":1760238513080,"version":"build-2065373602"},"reference-count":56,"publisher":"MDPI AG","issue":"8","license":[{"start":{"date-parts":[[2020,8,12]],"date-time":"2020-08-12T00:00:00Z","timestamp":1597190400000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100001700","name":"Ministry of Education, Culture, Sports, Science and Technology","doi-asserted-by":"publisher","award":["23117001, 23117005"],"award-info":[{"award-number":["23117001, 23117005"]}],"id":[{"id":"10.13039\/501100001700","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100001691","name":"Japan Society for the Promotion of Science","doi-asserted-by":"publisher","award":["17K19416; JP18H02650; JP20K16233"],"award-info":[{"award-number":["17K19416; JP18H02650; JP20K16233"]}],"id":[{"id":"10.13039\/501100001691","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Microorganisms"],"abstract":"<jats:p>Mitochondrial matrix proteins synthesized in the cytosol often contain amino (N)-terminal targeting sequences (NTSs), or alternately internal targeting sequences (ITSs), which enable them to be properly translocated to the organelle. Such sequences are also required for proteins targeted to mitochondrion-related organelles (MROs) that are present in a few species of anaerobic eukaryotes. Similar to other MROs, the mitosomes of the human intestinal parasite Entamoeba histolytica are highly degenerate, because a majority of the components involved in various processes occurring in the canonical mitochondria are either missing or modified. As of yet, sulfate activation continues to be the only identified role of the relic mitochondria of Entamoeba. Mitosomes influence the parasitic nature of E. histolytica, as the downstream cytosolic products of sulfate activation have been reported to be essential in proliferation and encystation. Here, we investigated the position of the targeting sequence of one of the mitosomal matrix enzymes involved in the sulfate activation pathway, ATP sulfurylase (AS). We confirmed by immunofluorescence assay and subcellular fractionation that hemagluttinin (HA)-tagged EhAS was targeted to mitosomes. However, its ortholog in the \u03b4-proteobacterium Desulfovibrio vulgaris, expressed as DvAS-HA in amoebic trophozoites, indicated cytosolic localization, suggesting a lack of recognizable mitosome targeting sequence in this protein. By expressing chimeric proteins containing swapped sequences between EhAS and DvAS in amoebic cells, we identified the ITSs responsible for mitosome targeting of EhAS. This observation is similar to other parasitic protozoans that harbor MROs, suggesting a convergent feature among various MROs in favoring ITS for the recognition and translocation of targeted proteins.<\/jats:p>","DOI":"10.3390\/microorganisms8081229","type":"journal-article","created":{"date-parts":[[2020,8,12]],"date-time":"2020-08-12T09:14:49Z","timestamp":1597223689000},"page":"1229","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":2,"title":["Import of Entamoeba histolytica Mitosomal ATP Sulfurylase Relies on Internal Targeting Sequences"],"prefix":"10.3390","volume":"8","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-4322-0145","authenticated-orcid":false,"given":"Herbert J.","family":"Santos","sequence":"first","affiliation":[{"name":"Department of Biomedical Chemistry, Graduate School of Medicine, The University of Tokyo, 7-3-1 Hongo, Bunkyo-ku, Tokyo 113-0033, Japan"},{"name":"Department of Parasitology, National Institute of Infectious Diseases, 1-23-1 Toyama, Shinjuku-ku, Tokyo 162-8640, Japan"},{"name":"Graduate School of Life and Environmental Sciences, University of Tsukuba, 1-1-1 Tennodai, Tsukuba, Ibaraki 305-8572, Japan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-2734-0836","authenticated-orcid":false,"given":"Yoko","family":"Chiba","sequence":"additional","affiliation":[{"name":"Department of Parasitology, National Institute of Infectious Diseases, 1-23-1 Toyama, Shinjuku-ku, Tokyo 162-8640, Japan"},{"name":"Graduate School of Life and Environmental Sciences, University of Tsukuba, 1-1-1 Tennodai, Tsukuba, Ibaraki 305-8572, Japan"},{"name":"Biofunctional Catalyst Research Team, RIKEN Center for Sustainable Resource Science (CSRS), Wako, Saitama 351-0198, Japan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Takashi","family":"Makiuchi","sequence":"additional","affiliation":[{"name":"Department of Parasitology, National Institute of Infectious Diseases, 1-23-1 Toyama, Shinjuku-ku, Tokyo 162-8640, Japan"},{"name":"Department of Infectious Diseases, Tokai University School of Medicine, 143 Shimokasuya, Isehara, Kanagawa 259-1193, Japan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Saki","family":"Arakawa","sequence":"additional","affiliation":[{"name":"Graduate School of Life and Environmental Sciences, University of Tsukuba, 1-1-1 Tennodai, Tsukuba, Ibaraki 305-8572, Japan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Yoshitaka","family":"Murakami","sequence":"additional","affiliation":[{"name":"Graduate School of Life and Environmental Sciences, University of Tsukuba, 1-1-1 Tennodai, Tsukuba, Ibaraki 305-8572, Japan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-4567-4768","authenticated-orcid":false,"given":"Kentaro","family":"Tomii","sequence":"additional","affiliation":[{"name":"Artificial Intelligence Research Center, National Institute of Advanced Industrial Science and Technology (AIST), 2-4-7 Aomi, Koto-ku, Tokyo 135-0064, Japan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-5664-0566","authenticated-orcid":false,"given":"Kenichiro","family":"Imai","sequence":"additional","affiliation":[{"name":"Cellular and Molecular Biotechnology Research Institute, National Institute of Advanced Industrial Science and Technology (AIST), 2-4-7 Aomi, Koto-ku, Tokyo 135-0064, Japan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-1354-5133","authenticated-orcid":false,"given":"Tomoyoshi","family":"Nozaki","sequence":"additional","affiliation":[{"name":"Department of Biomedical Chemistry, Graduate School of Medicine, The University of Tokyo, 7-3-1 Hongo, Bunkyo-ku, Tokyo 113-0033, Japan"},{"name":"Department of Parasitology, National Institute of Infectious Diseases, 1-23-1 Toyama, Shinjuku-ku, Tokyo 162-8640, Japan"},{"name":"Graduate School of Life and Environmental Sciences, University of Tsukuba, 1-1-1 Tennodai, Tsukuba, Ibaraki 305-8572, Japan"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2020,8,12]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"650","DOI":"10.1016\/j.tibs.2020.04.001","article-title":"Cytosolic Events in the Biogenesis of Mitochondrial Proteins","volume":"45","author":"Bykov","year":"2020","journal-title":"Trends Biochem. Sci."},{"key":"ref_2","doi-asserted-by":"crossref","unstructured":"Santos, H.J., Makiuchi, T., and Nozaki, T. (2018). Reinventing an Organelle: The Reduced Mitochondrion in Parasitic Protists. Trends Parasitol., 1\u201318.","DOI":"10.1016\/j.pt.2018.08.008"},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"314","DOI":"10.1126\/science.1127895","article-title":"Evolution of the molecular machines for protein import into mitochondria","volume":"313","author":"Dolezal","year":"2006","journal-title":"Science"},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"1113","DOI":"10.1074\/mcp.M114.043083","article-title":"MitoFates: Improved Prediction of Mitochondrial Targeting Sequences and Their Cleavage Sites","volume":"14","author":"Fukasawa","year":"2015","journal-title":"Mol. Cell. Proteom."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"1369","DOI":"10.1083\/jcb.201708044","article-title":"Tom70 enhances mitochondrial preprotein import efficiency by binding to internal targeting sequences","volume":"217","author":"Backes","year":"2018","journal-title":"J. Cell Biol."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"11752","DOI":"10.1073\/pnas.96.21.11752","article-title":"An internal targeting signal directing proteins into the mitochondrial intermembrane space","volume":"96","author":"Diekert","year":"1999","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"1574","DOI":"10.1093\/molbev\/msx096","article-title":"Origin and evolutionary alteration of the mitochondrial import system in eukaryotic lineages","volume":"34","author":"Fukasawa","year":"2017","journal-title":"Mol. Biol. Evol."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"605","DOI":"10.1038\/348605a0","article-title":"A yeast mitochondrial outer membrane protein essential for protein import and cell viability","volume":"348","author":"Baker","year":"1990","journal-title":"Nature"},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"516","DOI":"10.1038\/26780","article-title":"Tom40 forms the hydrophilic channel of the mitochondrial import pore for preproteins","volume":"395","author":"Hill","year":"1998","journal-title":"Nature"},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"395","DOI":"10.1038\/s41586-019-1680-7","article-title":"Structure of the mitochondrial import gate reveals distinct preprotein paths","volume":"575","author":"Araiso","year":"2019","journal-title":"Nature"},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"930","DOI":"10.1016\/j.bbamcr.2019.02.012","article-title":"Mitochondrial presequence import: Multiple regulatory knobs fine-tune mitochondrial biogenesis and homeostasis","volume":"1866","author":"Moulin","year":"2019","journal-title":"Biochim. Biophys. Acta Mol. Cell Res."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"3","DOI":"10.1016\/j.biochi.2013.11.018","article-title":"Highly divergent mitochondrion-related organelles in anaerobic parasitic protozoa","volume":"100","author":"Makiuchi","year":"2014","journal-title":"Biochimie"},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"409","DOI":"10.1146\/annurev.micro.62.081307.162826","article-title":"Mitochondrion-Related Organelles in Eukaryotic Protists","volume":"64","author":"Shiflett","year":"2010","journal-title":"Annu. Rev. Microbiol."},{"key":"ref_14","doi-asserted-by":"crossref","unstructured":"Dolezal, P., Dagley, M.J., Kono, M., Wolynec, P., Likic, V.A., Foo, J.H., Sedinova, M., Tachezy, J., Bachmann, A., and Bruchhaus, I. (2010). The Essentials of Protein Import in the Degenerate Mitochondrion of Entamoeba histolytica. PLoS Pathog., 6.","DOI":"10.1371\/journal.ppat.1000812"},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"628","DOI":"10.1016\/j.cell.2009.08.005","article-title":"Importing Mitochondrial Proteins: Machineries and Mechanisms","volume":"138","author":"Chacinska","year":"2009","journal-title":"Cell"},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"444","DOI":"10.1128\/MMBR.05024-11","article-title":"Biochemistry and Evolution of Anaerobic Energy Metabolism in Eukaryotes","volume":"76","author":"Mentel","year":"2012","journal-title":"Microbiol. Mol. Biol. Rev."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"21731","DOI":"10.1073\/pnas.0907106106","article-title":"Mitosomes in Entamoeba histolytica contain a sulfate activation pathway","volume":"106","author":"Yousuf","year":"2009","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"479","DOI":"10.1111\/mmi.13827","article-title":"Uniqueness of Entamoeba sulfur metabolism: Sulfolipid metabolism that plays pleiotropic roles in the parasitic life cycle","volume":"106","author":"Miyamoto","year":"2017","journal-title":"Mol. Microbiol."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"2095","DOI":"10.1016\/S0140-6736(12)61728-0","article-title":"Global and regional mortality from 235 causes of death for 20 age groups in 1990 and 2010: A systematic analysis for the Global Burden of Disease Study 2010","volume":"380","author":"Lozano","year":"2012","journal-title":"Lancet"},{"key":"ref_20","first-page":"E2884","article-title":"Entamoeba mitosomes play an important role in encystation by association with cholesteryl sulfate synthesis","volume":"112","author":"Miyamoto","year":"2015","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_21","first-page":"1","article-title":"Sulfate activation in mitosomes plays an important role in the proliferation of Entamoeba histolytica","volume":"5","author":"Makiuchi","year":"2011","journal-title":"PLoS Negl. Trop. Dis."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"1129","DOI":"10.1038\/srep01129","article-title":"Novel TPR-containing subunit of TOM complex functions as cytosolic receptor for Entamoeba mitosomal transport","volume":"3","author":"Makiuchi","year":"2013","journal-title":"Sci. Rep."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"1039","DOI":"10.1093\/molbev\/msu408","article-title":"Lateral gene transfer and gene duplication played a key role in the evolution of Mastigamoeba balamuthi hydrogenosomes","volume":"32","author":"Stairs","year":"2015","journal-title":"Mol. Biol. Evol."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"431","DOI":"10.1016\/0035-9203(78)90144-X","article-title":"A new medium for the axenic cultivation of Entamoeba histolytica and other Entamoeba","volume":"72","author":"Diamond","year":"1978","journal-title":"Trans. R. Soc. Trop. Med. Hyg."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1038\/srep08545","article-title":"A novel mitosomal \u03b2-barrel outer membrane protein in Entamoeba","volume":"5","author":"Santos","year":"2015","journal-title":"Sci. Rep."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"1471","DOI":"10.1111\/j.1462-5822.2009.01341.x","article-title":"Phosphatidylinositol-phosphates mediate cytoskeletal reorganization during phagocytosis via a unique modular protein consisting of RhoGEF\/DH and FYVE domains in the parasitic protozoon Entamoeba histolytica","volume":"11","author":"Okada","year":"2009","journal-title":"Cell. Microbiol."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"32445","DOI":"10.1074\/jbc.274.45.32445","article-title":"Characterization of the Gene Encoding Serine Acetyltransferase, a Regulated Enzyme of Cysteine Biosynthesis from the Protist Parasites Entamoeba histolytica and Entamoeba dispar","volume":"274","author":"Nozaki","year":"1999","journal-title":"J. Biol. Chem."},{"key":"ref_28","first-page":"23960","article-title":"Discovery of PPi-type Phosphoenolpyruvate Carboxykinase Genes in Eukaryotes and Bacteria","volume":"290","author":"Chiba","year":"2015","journal-title":"J. Biochem."},{"key":"ref_29","doi-asserted-by":"crossref","unstructured":"Santos, H.J., Hanadate, Y., Imai, K., and Nozaki, T. (2019). An Entamoeba-Specific Mitosomal Membrane Protein with Potential Association to the Golgi Apparatus. Genes, 10.","DOI":"10.3390\/genes10050367"},{"key":"ref_30","doi-asserted-by":"crossref","unstructured":"Tomii, K., Santos, H.J., and Nozaki, T. (2019). Genome-wide analysis of known and potential tetraspanins in Entamoeba histolytica. Genes, 10.","DOI":"10.3390\/genes10110885"},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"33","DOI":"10.1016\/S0166-6851(98)00129-7","article-title":"Molecular cloning and characterization of the genes encoding two isoforms of cysteine synthase in the enteric protozoan parasite Entamoeba histolytica","volume":"97","author":"Nozaki","year":"1998","journal-title":"Mol. Biochem. Parasitol."},{"key":"ref_32","first-page":"5","article-title":"Comparative protein structure modeling using MODELLER","volume":"56","author":"Webb","year":"2016","journal-title":"Curr. Protoc. Protein Sci."},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"6795","DOI":"10.1021\/bi010367w","article-title":"Crystal structure of ATP sulfurylase from Penicillium chrysogenum: Insights into the allosteric regulation of sulfate assimilation","volume":"40","author":"MacRae","year":"2001","journal-title":"Biochemistry"},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"594","DOI":"10.1093\/bioinformatics\/btg474","article-title":"FORTE: A profile-profile comparison tool for protein fold recognition","volume":"20","author":"Tomii","year":"2004","journal-title":"Bioinformatics"},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"1605","DOI":"10.1002\/jcc.20084","article-title":"UCSF Chimera\u2014A visualization system for exploratory research and analysis","volume":"25","author":"Pettersen","year":"2004","journal-title":"J. Comput. Chem."},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"4673","DOI":"10.1093\/nar\/22.22.4673","article-title":"CLUSTAL W: Improving the sensitivity of progressive multiple sequence alignment through sequence weighting, position-specific gap penalties and weight matrix choice","volume":"22","author":"Thompson","year":"1994","journal-title":"Nucleic Acids Res."},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"10","DOI":"10.1016\/j.exppara.2007.08.004","article-title":"Entamoeba histolytica mitosomes: Organelles in search of a function","volume":"118","author":"Aguilera","year":"2008","journal-title":"Exp. Parasitol."},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"926","DOI":"10.1128\/EC.00011-10","article-title":"Localization and targeting of an unusual pyridine nucleotide transhydrogenase in Entamoeba histolytica","volume":"9","author":"Yousuf","year":"2010","journal-title":"Eukaryot. Cell"},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"10924","DOI":"10.1073\/pnas.0500349102","article-title":"Giardia mitosomes and trichomonad hydrogenosomes share a common mode of protein targeting","volume":"102","author":"Dolezal","year":"2005","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"229","DOI":"10.1073\/pnas.95.1.229","article-title":"A mitochondrial-like chaperonin 60 gene in Giardia lamblia: Evidence that diplomonads once harbored an endosymbiont related to the progenitor of mitochondria","volume":"95","author":"Roger","year":"1998","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"30557","DOI":"10.1074\/jbc.M500787200","article-title":"Protein import, replication, and inheritance of a vestigial mitochondrion","volume":"280","author":"Regoes","year":"2005","journal-title":"J. Biol. Chem."},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"1750","DOI":"10.1128\/EC.00206-08","article-title":"Protein import into hydrogenosomes of Trichomonas vaginalis involves both N-terminal and internal targeting signals: A case study of thioredoxin reductases","volume":"7","author":"Mentel","year":"2008","journal-title":"Eukaryot. Cell"},{"key":"ref_43","doi-asserted-by":"crossref","first-page":"89","DOI":"10.1111\/jeu.12012","article-title":"The N-terminal sequences of four major hydrogenosomal proteins are not essential for import into hydrogenosomes of Trichomonas vaginalis","volume":"60","author":"Zimorski","year":"2013","journal-title":"J. Eukaryot. Microbiol."},{"key":"ref_44","doi-asserted-by":"crossref","first-page":"894","DOI":"10.1016\/j.tcb.2016.07.001","article-title":"The Role of Charge in Protein Targeting Evolution","volume":"26","author":"Garg","year":"2016","journal-title":"Trends Cell Biol."},{"key":"ref_45","doi-asserted-by":"crossref","unstructured":"\u0160m\u00edd, O., Matu\u0161kov\u00e1, A., Harris, S.R., Ku\u010dera, T., Novotn\u00fd, M., Horv\u00e1thov\u00e1, L., Hrd\u00fd, I., Kut\u011bjov\u00e1, E., Hirt, R.P., and Embley, T.M. (2008). Reductive evolution of the mitochondrial processing peptidases of the unicellular parasites Trichomonas vaginalis and Giardia intestinalis. PLoS Pathog., 4.","DOI":"10.1371\/journal.ppat.1000243"},{"key":"ref_46","doi-asserted-by":"crossref","first-page":"1501","DOI":"10.1016\/j.str.2014.07.015","article-title":"Molecular basis of the dynamic structure of the TIM23 complex in the mitochondrial intermembrane space","volume":"22","author":"Bajaj","year":"2014","journal-title":"Structure"},{"key":"ref_47","doi-asserted-by":"crossref","first-page":"1154","DOI":"10.1111\/j.1365-2958.2007.05719.x","article-title":"A functionally divergent hydrogenosomal peptidase with protomitochondrial ancestry","volume":"64","author":"Brown","year":"2007","journal-title":"Mol. Microbiol."},{"key":"ref_48","doi-asserted-by":"crossref","first-page":"113","DOI":"10.1093\/molbev\/msr165","article-title":"A small Tim homohexamer in the relict mitochondrion of Cryptosporidium","volume":"29","author":"Alcock","year":"2012","journal-title":"Mol. Biol. Evol."},{"key":"ref_49","doi-asserted-by":"crossref","first-page":"441","DOI":"10.1126\/science.1094786","article-title":"Complete genome sequence of the apicomplexan, Cryptosporidium parvum","volume":"304","author":"Abrahamsen","year":"2004","journal-title":"Science"},{"key":"ref_50","doi-asserted-by":"crossref","unstructured":"Denoeud, F., Roussel, M., Noel, B., Wawrzyniak, I., Da Silva, C., Diogon, M., Viscogliosi, E., Brochier-Armanet, C., Couloux, A., and Poulain, J. (2011). Genome sequence of the stramenopile Blastocystis, a human anaerobic parasite. Genome Biol., 12.","DOI":"10.1186\/gb-2011-12-3-r29"},{"key":"ref_51","doi-asserted-by":"crossref","first-page":"450","DOI":"10.1038\/35106579","article-title":"Genome sequence and gene compaction of the eukaryote parasite Encephalitozoon cuniculi","volume":"414","author":"Katinka","year":"2001","journal-title":"Nature"},{"key":"ref_52","doi-asserted-by":"crossref","first-page":"15916","DOI":"10.1073\/pnas.0604109103","article-title":"Microsporidian mitosomes retain elements of the general mitochondrial targeting system","volume":"103","author":"Burri","year":"2006","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_53","first-page":"1","article-title":"Hydrogenosomes in the diplomonad Spironucleus salmonicida","volume":"4","author":"Einarsson","year":"2013","journal-title":"Nat. Commun."},{"key":"ref_54","doi-asserted-by":"crossref","first-page":"2813","DOI":"10.1093\/gbe\/evy215","article-title":"A single tim translocase in themitosomes of Giardia intestinalis illustrates convergence of protein import machines in anaerobic eukaryotes","volume":"10","author":"Voleman","year":"2018","journal-title":"Genome Biol. Evol."},{"key":"ref_55","doi-asserted-by":"crossref","first-page":"10","DOI":"10.1016\/j.molbiopara.2016.01.001","article-title":"Screening and discovery of lineage-specific mitosomal membrane proteins in Entamoeba histolytica","volume":"209","author":"Santos","year":"2016","journal-title":"Mol. Biochem. Parasitol."},{"key":"ref_56","doi-asserted-by":"crossref","first-page":"2716","DOI":"10.1093\/gbe\/evv175","article-title":"Conservation of transit peptide-independent protein import into the mitochondrial and hydrogenosomal matrix","volume":"7","author":"Garg","year":"2015","journal-title":"Genome Biol. Evol."}],"container-title":["Microorganisms"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2076-2607\/8\/8\/1229\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,11]],"date-time":"2025-10-11T09:59:42Z","timestamp":1760176782000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2076-2607\/8\/8\/1229"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2020,8,12]]},"references-count":56,"journal-issue":{"issue":"8","published-online":{"date-parts":[[2020,8]]}},"alternative-id":["microorganisms8081229"],"URL":"https:\/\/doi.org\/10.3390\/microorganisms8081229","relation":{},"ISSN":["2076-2607"],"issn-type":[{"type":"electronic","value":"2076-2607"}],"subject":[],"published":{"date-parts":[[2020,8,12]]}}}