{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,10,10]],"date-time":"2025-10-10T01:38:35Z","timestamp":1760060315900,"version":"build-2065373602"},"reference-count":52,"publisher":"MDPI AG","issue":"8","license":[{"start":{"date-parts":[[2025,8,19]],"date-time":"2025-08-19T00:00:00Z","timestamp":1755561600000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100004837","name":"\u201cMinisterio de Ciencia e Innovaci\u00f3n\u201d from Spain","doi-asserted-by":"publisher","award":["PID2023-147765OB-C21, -C22"],"award-info":[{"award-number":["PID2023-147765OB-C21, -C22"]}],"id":[{"id":"10.13039\/501100004837","id-type":"DOI","asserted-by":"publisher"}]},{"name":"Fundaci\u00f3n Roviralta (Chair \u201cMaria Francisca de Roviralta of Molecular Parasitology, Leishmaniasis and One Health\u201d)","award":["PID2023-147765OB-C21, -C22"],"award-info":[{"award-number":["PID2023-147765OB-C21, -C22"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Biomolecules"],"abstract":"<jats:p>Leishmaniasis is caused by protozoan parasites from the genus Leishmania and remains one of the major threats to global health, impacting millions of people worldwide as well as animals including dogs. Several treatments have been used for managing leishmaniasis; nevertheless, drug resistance has emerged as an important obstacle to disease control. Therefore, there is an urgent need to discover new therapeutic targets. The aim of this work was to study the role played by the breast cancer associated 1 C-terminal (BRCT) domain from LmjPES protein (Pescadillo ribosomal biogenesis factor) in Leishmania major\u2018s pathogenesis through the construction of novel genomic tools. For this purpose, Leishmania integrative plasmids that were able to express the BRCT domain from LmjPES and a hypothetical defective LmjPES lacking this BRCT domain were constructed. It was observed that the overexpression of the aforementioned BRCT domain in L. major dysregulated the mRNA expression of 152 genes (95 up-regulated and 57 down-regulated) in respect to control parasites. Furthermore, clustering studies of these altered genes revealed an enrichment in genes related to metabolic processes, transporter activity, response to stimuli, and protein folding, which are categories described to be associated with the metacyclogenesis process and parasite survival. Interestingly, these genes reached normal levels of expression in parasites transfected with a defective LmjPES (a mutated gene lacking the coding sequence of the BRCT domain). In addition, it was found that the footpad of mice inoculated with LmjPES BRCT-overexpressing parasites had significantly greater inflammation compared to the size of the footpad of animals infected with the control parasites. Based on all these results, it was suggested that the BRCT domain from LmjPES might play a role in L. major\u2018s infection process and pathogenesis.<\/jats:p>","DOI":"10.3390\/biom15081191","type":"journal-article","created":{"date-parts":[[2025,8,19]],"date-time":"2025-08-19T10:59:01Z","timestamp":1755601141000},"page":"1191","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["The Role of BRCT Domain from LmjPES in Leishmania\u00a0major Pathogenesis"],"prefix":"10.3390","volume":"15","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-1443-6513","authenticated-orcid":false,"given":"Esther","family":"Larrea","sequence":"first","affiliation":[{"name":"Unit of Translational Medicine, IdiSNA (Navarra Institute for Health Research), University of Navarra, 31009 Pamplona, Navarra, Spain"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-9183-6369","authenticated-orcid":false,"given":"Jos\u00e9","family":"Pe\u00f1a-Guerrero","sequence":"additional","affiliation":[{"name":"Department of Microbiology and Parasitology, IdiSNA (Navarra Institute for Health Research), University of Navarra, 31009 Pamplona, Navarra, Spain"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-5254-8121","authenticated-orcid":false,"given":"Celia","family":"Fern\u00e1ndez-Rubio","sequence":"additional","affiliation":[{"name":"Department of Microbiology and Parasitology, IdiSNA (Navarra Institute for Health Research), University of Navarra, 31009 Pamplona, Navarra, Spain"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-2167-092X","authenticated-orcid":false,"given":"Aroia","family":"Burguete-Mikeo","sequence":"additional","affiliation":[{"name":"Department of Microbiology and Parasitology, IdiSNA (Navarra Institute for Health Research), University of Navarra, 31009 Pamplona, Navarra, Spain"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-0547-681X","authenticated-orcid":false,"given":"Elizabeth","family":"Guruceaga","sequence":"additional","affiliation":[{"name":"Bioinformatics Platform, Center for Applied Medical Research, IdiSNA (Navarra Institute for Health Research), University of Navarra, 31009 Pamplona, Navarra, Spain"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-2193-7316","authenticated-orcid":false,"given":"Paul","family":"Nguewa","sequence":"additional","affiliation":[{"name":"Department of Microbiology and Parasitology, IdiSNA (Navarra Institute for Health Research), University of Navarra, 31009 Pamplona, Navarra, Spain"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2025,8,19]]},"reference":[{"key":"ref_1","unstructured":"(2025, February 24). Leishmaniasis. Available online: https:\/\/www.who.int\/news-room\/fact-sheets\/detail\/leishmaniasis."},{"key":"ref_2","doi-asserted-by":"crossref","unstructured":"Burguete-Mikeo, A., Fern\u00e1ndez-Rubio, C., Pe\u00f1a-Guerrero, J., El-Dirany, R., Gainza, L., Carasa Buj, B., and Nguewa, P.A. (2023). Characterization of Leishmania parasites isolated from naturally infected mammals. Animals, 13.","DOI":"10.3390\/ani13132153"},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"976","DOI":"10.1016\/j.pt.2021.07.003","article-title":"Leishmaniasis: The act of transmission","volume":"76","author":"Serafim","year":"2021","journal-title":"Trends Parasitol."},{"key":"ref_4","doi-asserted-by":"crossref","unstructured":"Ranatunga, M., Deacon, A., Harbige, L.S., Dyer, P., Boateng, J., and Getti, G.T.M. (2024). Ex vivo analysis of the association of GFP-expressing L. aethiopica and L. mexicana with human peripheral blood-derived (PBD) leukocytes over 24 hours. Microorganisms, 12.","DOI":"10.3390\/microorganisms12091909"},{"key":"ref_5","doi-asserted-by":"crossref","unstructured":"dos Santos Meira, C., and Gedamu, L. (2019). Protective or detrimental? understanding the role of host immunity in leishmaniasis. Microorganisms, 7.","DOI":"10.3390\/microorganisms7120695"},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"969","DOI":"10.4049\/jimmunol.165.2.969","article-title":"A natural model of Leishmania major infection reveals a prolonged \u201cSilent\u201d phase of parasite amplification in the skin before the onset of lesion formation and immunity","volume":"165","author":"Belkaid","year":"2000","journal-title":"J. Immunol."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"239","DOI":"10.1111\/j.1574-695X.2009.00607.x","article-title":"Correlation of parasitic load with interleukin-4 response in patients with cutaneous leishmaniasis due to Leishmania tropica","volume":"57","author":"Kumar","year":"2009","journal-title":"FEMS Immunol. Med. Microbiol."},{"key":"ref_8","doi-asserted-by":"crossref","unstructured":"Hanau, S., Maritati, M., Contini, C., Trentini, A., Manfrinato, M.C., and Almugadam, S.H. (2025). Commentary on the issue of Leishmania infection: Focus on some pathogenetic, clinical, and epidemiological aspects. Vet. Sci., 12.","DOI":"10.3390\/vetsci12060536"},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"108400","DOI":"10.1016\/j.intimp.2021.108400","article-title":"Exploring the paradox of defense between host and Leishmania parasite","volume":"102","author":"Kumari","year":"2022","journal-title":"Int. Immunopharmacol."},{"key":"ref_10","doi-asserted-by":"crossref","unstructured":"Karamysheva, Z.N., Gutierrez Guarnizo, S.A., and Karamyshev, A.L. (2020). Regulation of translation in the protozoan parasite Leishmania. Int. J. Mol. Sci., 21.","DOI":"10.3390\/ijms21082981"},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"251","DOI":"10.1016\/j.pt.2023.01.012","article-title":"Drug resistance in Leishmania: Does it really matter?","volume":"39","author":"Domagalska","year":"2023","journal-title":"Trends Parasitol."},{"key":"ref_12","doi-asserted-by":"crossref","unstructured":"Cosma, C., Maia, C., Khan, N., Infantino, M., and Del Riccio, M. (2024). Leishmaniasis in humans and animals: A one health approach for surveillance, prevention and control in a changing world. Trop. Med. Infect. Dis., 9.","DOI":"10.3390\/tropicalmed9110258"},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"436","DOI":"10.1126\/science.1112680","article-title":"The genome of the kinetoplastid parasite, Leishmania major","volume":"309","author":"Ivens","year":"2005","journal-title":"Science"},{"key":"ref_14","doi-asserted-by":"crossref","unstructured":"Algarabel, M., Fern\u00e1ndez-rubio, C., Musilova, K., Pe\u00f1a-guerrero, J., Vacas, A., Larrea, E., and Nguewa, P.A. (2021). In Leishmania major, the homolog of the oncogene Pes1 may play a critical role in parasite infectivity. Int. J. Mol. Sci., 22.","DOI":"10.3390\/ijms222212592"},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"48896","DOI":"10.1016\/j.gene.2024.148896","article-title":"Emerging role of PES1 in disease: A promising therapeutic target?","volume":"932","author":"Yuan","year":"2025","journal-title":"Gene"},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1016\/j.biocel.2018.08.014","article-title":"PES1 Is Transcriptionally regulated by BRD4 and promotes cell proliferation and glycolysis in hepatocellular carcinoma","volume":"104","author":"Fan","year":"2018","journal-title":"Int. J. Biochem. Cell Biol."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"268","DOI":"10.7150\/jca.58982","article-title":"The functional role of Pescadillo ribosomal biogenesis factor 1 in cancer","volume":"13","author":"Li","year":"2022","journal-title":"J. Cancer"},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"6656","DOI":"10.1074\/jbc.M008536200","article-title":"Pescadillo, a novel cell cycle regulatory protein abnormally expressed in malignant cells","volume":"276","author":"Kinoshita","year":"2001","journal-title":"J. Biol. Chem."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"2765","DOI":"10.1016\/j.molcel.2021.05.010","article-title":"Structural insight into BRCA1-BARD1 complex recruitment to damaged chromatin","volume":"81","author":"Dai","year":"2021","journal-title":"Mol. Cell"},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"89","DOI":"10.1007\/s10689-005-2579-z","article-title":"Insights into the molecular basis of human hereditary breast cancer from studies of the BRCA1 BRCT domain","volume":"5","author":"Glover","year":"2006","journal-title":"Fam. Cancer"},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"275","DOI":"10.2174\/138920208784533610","article-title":"NBS1 heterozygosity and cancer Risk","volume":"9","author":"Masi","year":"2008","journal-title":"Curr. Genom."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"789","DOI":"10.1093\/nar\/gkl1058","article-title":"The BRCT domain of mammalian Pes1 is crucial for nucleolar localization and rRNA processing","volume":"35","author":"Grimm","year":"2007","journal-title":"Nucleic Acids Res."},{"key":"ref_23","doi-asserted-by":"crossref","unstructured":"Pe\u00f1a-Guerrero, J., Fern\u00e1ndez-Rubio, C., Burguete-Mikeo, A., El-Dirany, R., Garc\u00eda-Sosa, A.T., and Nguewa, P. (2021). Discovery and validation of Lmj_04_BRCT domain, a novel therapeutic target: Identification of candidate drugs for leishmaniasis. Int. J. Mol. Sci., 22.","DOI":"10.3390\/ijms221910493"},{"key":"ref_24","doi-asserted-by":"crossref","unstructured":"Larrea, E., Fern\u00e1ndez-Rubio, C., Pe\u00f1a-Guerrero, J., Guruceaga, E., and Nguewa, P.A. (2022). The BRCT domain from the homologue of the oncogene PES1 in Leishmania major (LmjPES) promotes malignancy and drug resistance in mammalian cells. Int. J. Mol. Sci., 23.","DOI":"10.3390\/ijms232113203"},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"327","DOI":"10.1016\/0166-6851(93)90231-L","article-title":"Rapid Isolation of DNA from Trypanosomatid protozoa using a simple \u201cmini-Prep\u201d procedure","volume":"59","author":"Cross","year":"1993","journal-title":"Mol. Biochem. Parasitol."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"564","DOI":"10.4049\/jimmunol.135.1.564","article-title":"Identification of cell surface carbohydrate and antigenic changes between noninfective and infective developmental stages of Leishmania major promastigotes","volume":"135","author":"Sacks","year":"1985","journal-title":"J. Immunol."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"217","DOI":"10.1016\/S0166-6851(03)00079-3","article-title":"Improvements in transfection efficiency and tests of RNA interference (RNAi) approaches in the protozoan parasite Leishmania","volume":"128","author":"Robinson","year":"2003","journal-title":"Mol. Biochem. Parasitol."},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"2114","DOI":"10.1093\/bioinformatics\/btu170","article-title":"Trimmomatic: A flexible trimmer for Illumina sequence data","volume":"30","author":"Bolger","year":"2014","journal-title":"Bioinformatics"},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"15","DOI":"10.1093\/bioinformatics\/bts635","article-title":"STAR: Ultrafast universal RNA-Seq aligner","volume":"29","author":"Dobin","year":"2013","journal-title":"Bioinformatics"},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"923","DOI":"10.1093\/bioinformatics\/btt656","article-title":"FeatureCounts: An efficient general purpose program for assigning sequence reads to genomic features","volume":"30","author":"Liao","year":"2014","journal-title":"Bioinformatics"},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"D996","DOI":"10.1093\/nar\/gkab1007","article-title":"Ensembl Genomes 2022: An expanding genome resource for non-vertebrates","volume":"50","author":"Yates","year":"2022","journal-title":"Nucleic Acids Res."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"275","DOI":"10.1007\/978-1-4939-8757-3_16","article-title":"Cluster analysis of untargeted metabolomic experiments","volume":"1859","author":"Heinemann","year":"2019","journal-title":"Methods Mol. Biol."},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"139","DOI":"10.1093\/bioinformatics\/btp616","article-title":"EdgeR: A bioconductor package for differential expression analysis of digital gene expression data","volume":"26","author":"Robinson","year":"2010","journal-title":"Bioinformatics"},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"e47","DOI":"10.1093\/nar\/gkv007","article-title":"Limma powers differential expression analyses for RNA-sequencing and microarray studies","volume":"43","author":"Ritchie","year":"2015","journal-title":"Nucleic Acids Res."},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"284","DOI":"10.1089\/omi.2011.0118","article-title":"ClusterProfiler: An R package for comparing biological themes among gene clusters","volume":"16","author":"Yu","year":"2012","journal-title":"OMICS"},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"D587","DOI":"10.1093\/nar\/gkac963","article-title":"KEGG for taxonomy-based analysis of pathways and genomes","volume":"51","author":"Kanehisa","year":"2022","journal-title":"Nucleic Acids Res."},{"key":"ref_37","doi-asserted-by":"crossref","unstructured":"Vacas, A., Fern\u00e1ndez-Rubio, C., Larrea, E., Pe\u00f1a-Guerrero, J., and Nguewa, P.A. (2020). LmjF.22.0810 from Leishmania major modulates the Th2-type immune response and is involved in leishmaniasis outcome. Biomedicines, 8.","DOI":"10.3390\/biomedicines8110452"},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"104557","DOI":"10.1016\/j.micpath.2020.104557","article-title":"Quantitative proteomic analysis reveals differentially expressed proteins in Leishmania major metacyclogenesis","volume":"149","author":"Zali","year":"2020","journal-title":"Microb. Pathog."},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1016\/j.jgar.2019.01.023","article-title":"Drug resistance in protozoan parasites: An incessant wrestle for survival","volume":"18","author":"Pramanik","year":"2019","journal-title":"J. Glob. Antimicrob. Resist."},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"637","DOI":"10.1016\/j.exppara.2010.12.006","article-title":"Fluorescent Leishmania species: Development of stable GFP expression and its application for in vitro and in vivo studies","volume":"127","author":"Bolhassani","year":"2011","journal-title":"Exp. Parasitol."},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"251","DOI":"10.1016\/S0166-6851(00)00195-X","article-title":"Targeted integration into a rRNA locus results in uniform and high level expression of transgenes in Leishmania amastigotes","volume":"107","author":"Mottram","year":"2000","journal-title":"Mol. Biochem. Parasitol."},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"287","DOI":"10.1016\/j.exppara.2012.12.007","article-title":"Release of the cytosolic tryparedoxin peroxidase into the incubation medium and a different profile of cytosolic and mitochondrial peroxiredoxin expression in H2O2-treated Trypanosoma cruzi tissue culture-derived trypomastigotes","volume":"133","author":"Gadelha","year":"2013","journal-title":"Exp. Parasitol."},{"key":"ref_43","doi-asserted-by":"crossref","first-page":"1860","DOI":"10.1021\/pr400923g","article-title":"Identification of virulence factors in Leishmania infantum strains by a proteomic approach","volume":"13","author":"Fialho","year":"2014","journal-title":"J. Proteome Res."},{"key":"ref_44","doi-asserted-by":"crossref","first-page":"25","DOI":"10.1007\/s10123-022-00259-4","article-title":"An insight into differential protein abundance throughout Leishmania donovani promastigote growth and differentiation","volume":"26","author":"Alcolea","year":"2023","journal-title":"Int. Microbiol."},{"key":"ref_45","doi-asserted-by":"crossref","unstructured":"Clos, J., Gr\u00fcnebast, J., and Holm, M. (2022). Promastigote-to-amastigote conversion in Leishmania spp.-A molecular view. Pathogens, 11.","DOI":"10.3390\/pathogens11091052"},{"key":"ref_46","doi-asserted-by":"crossref","first-page":"78","DOI":"10.1186\/s13568-022-01419-5","article-title":"The role of ATP-binding cassette transporter genes expression in treatment failure cutaneous leishmaniasis","volume":"12","author":"Boozhmehrani","year":"2022","journal-title":"AMB Express"},{"key":"ref_47","doi-asserted-by":"crossref","unstructured":"Roy, G., Bhattacharya, A., Leprohon, P., and Ouellette, M. (2021). Decreased glutamate transport in acivicin resistant Leishmania tarentolae. PLoS Negl. Trop. Dis., 15.","DOI":"10.1371\/journal.pntd.0010046"},{"key":"ref_48","doi-asserted-by":"crossref","first-page":"1713","DOI":"10.1128\/EC.00152-06","article-title":"Modulation of Leishmania ABC protein gene expression through life stages and among drug-resistant parasites","volume":"5","author":"Leprohon","year":"2006","journal-title":"Eukaryot. Cell"},{"key":"ref_49","doi-asserted-by":"crossref","first-page":"209","DOI":"10.1111\/j.1574-695X.1998.tb01129.x","article-title":"The Leishmania promastigote surface antigen-2 (PSA-2) is specifically recognised by Th1 cells in humans with naturally acquired immunity to L. major","volume":"20","author":"Kemp","year":"1998","journal-title":"FEMS Immunol. Med. Microbiol."},{"key":"ref_50","doi-asserted-by":"crossref","first-page":"1175","DOI":"10.1111\/j.1365-2958.2010.07130.x","article-title":"Overexpression of a single Leishmania major gene enhances parasite infectivity in vivo and in vitro","volume":"76","author":"Reiling","year":"2010","journal-title":"Mol. Microbiol."},{"key":"ref_51","doi-asserted-by":"crossref","first-page":"1457","DOI":"10.1111\/j.1365-2958.2006.05205.x","article-title":"Leishmania histone H1 overexpression delays parasite cell-cycle progression, parasite differentiation and reduces Leishmania infectivity in vivo","volume":"60","author":"Smirlis","year":"2006","journal-title":"Mol. Microbiol."},{"key":"ref_52","doi-asserted-by":"crossref","first-page":"511","DOI":"10.1046\/j.1462-5822.2001.00135.x","article-title":"Phenotypic changes associated with deletion and overexpression of a stage-regulated gene family in Leishmania","volume":"3","author":"Mckean","year":"2001","journal-title":"Cell. Microbiol."}],"container-title":["Biomolecules"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2218-273X\/15\/8\/1191\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,9]],"date-time":"2025-10-09T18:30:48Z","timestamp":1760034648000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2218-273X\/15\/8\/1191"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2025,8,19]]},"references-count":52,"journal-issue":{"issue":"8","published-online":{"date-parts":[[2025,8]]}},"alternative-id":["biom15081191"],"URL":"https:\/\/doi.org\/10.3390\/biom15081191","relation":{},"ISSN":["2218-273X"],"issn-type":[{"type":"electronic","value":"2218-273X"}],"subject":[],"published":{"date-parts":[[2025,8,19]]}}}