Differential composition of the translational machinery of Trypanosoma cruzi during metacyclogenesis and cell cycle
Resumen:
Trypanosoma cruzi (T. cruzi), the causative agent of Chagas disease, primarily controls gene expression through post-transcriptional mechanisms. Our research group has identified translational regulation as a key process occurring during both metacyclogenesis and the G1/S transition of the cell cycle. Recent findings using various approaches suggest that ribosomes may exhibit compositional diversity at the protein level, potentially influencing regulatory outcomes. Based on this, we aimed to perform an in-depth characterization of the translational machinery in T. cruzi, focusing on the epimastigote and metacyclic trypomastigote stages of the life cycle, as well as on parasites synchronized in G1 and S phases. As a first step, we curated the current annotation of ribosomal proteins (RPs), assessing copy number, ribosomal location, N- and C-terminal extensions, expression levels, and possible extra-ribosomal roles. Experimentally, Ribo-Seq analysis revealed a global repression of RP mRNA translation in the metacyclic trypomastigote stage. However, some RP transcripts showed distinct behavior, suggesting selective resistance to this repression. Additionally, we applied a multi-omics approach—combining RNA-Seq, Ribo-Seq, and proteomics—on parasites synchronized in G1 and S phases. This analysis uncovered individual differences in translational efficiency of RP mRNAs, as well as in the steady-state protein levels of RPs. These findings support the idea that the composition of the translational machinery may vary during these biological transitions. To further explore this hypothesis, we conducted quantitative proteomic analysis of ribosome-enriched fractions obtained from epimastigotes, metacyclic trypomastigotes, and cell cycle–synchronized parasites in G1 and S phases. Through these experiments, we found that certain ribosomal proteins exhibit differential abundance in ribosome-enriched fractions across the different life cycle stages. We also observed variations in the levels of proteins associated with ribosomes. The differential abundance of components of the translational machinery could have regulatory implications, representing a potential new layer of gene expression regulation in T. cruzi.
| 2025 | |
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Trypanosoma cruzi Ribosoma Regulación de la expresión génica Ciencias Naturales y Exactas Ciencias Biológicas Bioquímica y Biología Molecular |
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| Inglés | |
| Instituto de Investigaciones Biológicas Clemente Estable | |
| IIBCE en REDI | |
| https://hdl.handle.net/20.500.12381/5357 | |
| Acceso abierto | |
| Reconocimiento 4.0 Internacional. (CC BY) |
| _version_ | 1880371848932229120 |
|---|---|
| author | Rivara, Martín |
| author2 | Radío, Santiago Chávez, Santiago Duhagon, María Ana Smircich, Pablo Sotelo, José |
| author2_role | author author author author author |
| author_facet | Rivara, Martín Radío, Santiago Chávez, Santiago Duhagon, María Ana Smircich, Pablo Sotelo, José |
| author_role | author |
| bitstream.checksum.fl_str_mv | fcc8b2ddb26c820405a37f9e0cae7f18 e57a516e558144ff4ccd80752c8061b2 |
| bitstream.checksumAlgorithm.fl_str_mv | MD5 MD5 |
| bitstream.url.fl_str_mv | https://redi.anii.org.uy/jspui/bitstream/20.500.12381/5357/2/license.txt https://redi.anii.org.uy/jspui/bitstream/20.500.12381/5357/1/Abstract%20MolBio%202025%20RIVARA.pdf |
| collection | IIBCE en REDI |
| dc.creator.none.fl_str_mv | Rivara, Martín Radío, Santiago Chávez, Santiago Duhagon, María Ana Smircich, Pablo Sotelo, José |
| dc.date.accessioned.none.fl_str_mv | 2026-01-12T18:27:05Z |
| dc.date.available.none.fl_str_mv | 2026-01-12T18:27:05Z |
| dc.date.issued.none.fl_str_mv | 2025-09-29 |
| dc.description.abstract.none.fl_txt_mv | Trypanosoma cruzi (T. cruzi), the causative agent of Chagas disease, primarily controls gene expression through post-transcriptional mechanisms. Our research group has identified translational regulation as a key process occurring during both metacyclogenesis and the G1/S transition of the cell cycle. Recent findings using various approaches suggest that ribosomes may exhibit compositional diversity at the protein level, potentially influencing regulatory outcomes. Based on this, we aimed to perform an in-depth characterization of the translational machinery in T. cruzi, focusing on the epimastigote and metacyclic trypomastigote stages of the life cycle, as well as on parasites synchronized in G1 and S phases. As a first step, we curated the current annotation of ribosomal proteins (RPs), assessing copy number, ribosomal location, N- and C-terminal extensions, expression levels, and possible extra-ribosomal roles. Experimentally, Ribo-Seq analysis revealed a global repression of RP mRNA translation in the metacyclic trypomastigote stage. However, some RP transcripts showed distinct behavior, suggesting selective resistance to this repression. Additionally, we applied a multi-omics approach—combining RNA-Seq, Ribo-Seq, and proteomics—on parasites synchronized in G1 and S phases. This analysis uncovered individual differences in translational efficiency of RP mRNAs, as well as in the steady-state protein levels of RPs. These findings support the idea that the composition of the translational machinery may vary during these biological transitions. To further explore this hypothesis, we conducted quantitative proteomic analysis of ribosome-enriched fractions obtained from epimastigotes, metacyclic trypomastigotes, and cell cycle–synchronized parasites in G1 and S phases. Through these experiments, we found that certain ribosomal proteins exhibit differential abundance in ribosome-enriched fractions across the different life cycle stages. We also observed variations in the levels of proteins associated with ribosomes. The differential abundance of components of the translational machinery could have regulatory implications, representing a potential new layer of gene expression regulation in T. cruzi. |
| dc.identifier.anii.es.fl_str_mv | POS_NAC_2021_1_170006 |
| dc.identifier.uri.none.fl_str_mv | https://hdl.handle.net/20.500.12381/5357 |
| dc.language.iso.none.fl_str_mv | eng |
| dc.rights.*.fl_str_mv | Acceso abierto |
| dc.rights.license.none.fl_str_mv | Reconocimiento 4.0 Internacional. (CC BY) |
| dc.rights.none.fl_str_mv | info:eu-repo/semantics/openAccess |
| dc.source.es.fl_str_mv | 4th Molecular Biosystems Conference: Eukaryotic Gene Regulation & Functional Genomics, Puerto Varas, 09/2025 |
| dc.source.none.fl_str_mv | reponame:IIBCE en REDI instname:Instituto de Investigaciones Biológicas Clemente Estable instacron:Instituto de Investigaciones Biológicas Clemente Estable |
| dc.subject.anii.none.fl_str_mv | Ciencias Naturales y Exactas Ciencias Biológicas Bioquímica y Biología Molecular |
| dc.subject.es.fl_str_mv | Trypanosoma cruzi Ribosoma Regulación de la expresión génica |
| dc.title.none.fl_str_mv | Differential composition of the translational machinery of Trypanosoma cruzi during metacyclogenesis and cell cycle |
| dc.type.es.fl_str_mv | Documento de conferencia |
| dc.type.none.fl_str_mv | info:eu-repo/semantics/conferenceObject |
| dc.type.version.es.fl_str_mv | Publicado |
| dc.type.version.none.fl_str_mv | info:eu-repo/semantics/publishedVersion |
| description | Trypanosoma cruzi (T. cruzi), the causative agent of Chagas disease, primarily controls gene expression through post-transcriptional mechanisms. Our research group has identified translational regulation as a key process occurring during both metacyclogenesis and the G1/S transition of the cell cycle. Recent findings using various approaches suggest that ribosomes may exhibit compositional diversity at the protein level, potentially influencing regulatory outcomes. Based on this, we aimed to perform an in-depth characterization of the translational machinery in T. cruzi, focusing on the epimastigote and metacyclic trypomastigote stages of the life cycle, as well as on parasites synchronized in G1 and S phases. As a first step, we curated the current annotation of ribosomal proteins (RPs), assessing copy number, ribosomal location, N- and C-terminal extensions, expression levels, and possible extra-ribosomal roles. Experimentally, Ribo-Seq analysis revealed a global repression of RP mRNA translation in the metacyclic trypomastigote stage. However, some RP transcripts showed distinct behavior, suggesting selective resistance to this repression. Additionally, we applied a multi-omics approach—combining RNA-Seq, Ribo-Seq, and proteomics—on parasites synchronized in G1 and S phases. This analysis uncovered individual differences in translational efficiency of RP mRNAs, as well as in the steady-state protein levels of RPs. These findings support the idea that the composition of the translational machinery may vary during these biological transitions. To further explore this hypothesis, we conducted quantitative proteomic analysis of ribosome-enriched fractions obtained from epimastigotes, metacyclic trypomastigotes, and cell cycle–synchronized parasites in G1 and S phases. Through these experiments, we found that certain ribosomal proteins exhibit differential abundance in ribosome-enriched fractions across the different life cycle stages. We also observed variations in the levels of proteins associated with ribosomes. The differential abundance of components of the translational machinery could have regulatory implications, representing a potential new layer of gene expression regulation in T. cruzi. |
| eu_rights_str_mv | openAccess |
| format | conferenceObject |
| id | IIBCE_88f26844ca391317f8e4ffeb4602e849 |
| identifier_str_mv | POS_NAC_2021_1_170006 |
| instacron_str | Instituto de Investigaciones Biológicas Clemente Estable |
| institution | Instituto de Investigaciones Biológicas Clemente Estable |
| instname_str | Instituto de Investigaciones Biológicas Clemente Estable |
| language | eng |
| network_acronym_str | IIBCE |
| network_name_str | IIBCE en REDI |
| oai_identifier_str | oai:redi.anii.org.uy:20.500.12381/5357 |
| publishDate | 2025 |
| reponame_str | IIBCE en REDI |
| repository.mail.fl_str_mv | vcarballo@iibce.edu.uy |
| repository.name.fl_str_mv | IIBCE en REDI - Instituto de Investigaciones Biológicas Clemente Estable |
| repository_id_str | 9421_3 |
| rights_invalid_str_mv | Reconocimiento 4.0 Internacional. (CC BY) Acceso abierto |
| spelling | Reconocimiento 4.0 Internacional. (CC BY)Acceso abiertoinfo:eu-repo/semantics/openAccess2026-01-12T18:27:05Z2026-01-12T18:27:05Z2025-09-29https://hdl.handle.net/20.500.12381/5357POS_NAC_2021_1_170006Trypanosoma cruzi (T. cruzi), the causative agent of Chagas disease, primarily controls gene expression through post-transcriptional mechanisms. Our research group has identified translational regulation as a key process occurring during both metacyclogenesis and the G1/S transition of the cell cycle. Recent findings using various approaches suggest that ribosomes may exhibit compositional diversity at the protein level, potentially influencing regulatory outcomes. Based on this, we aimed to perform an in-depth characterization of the translational machinery in T. cruzi, focusing on the epimastigote and metacyclic trypomastigote stages of the life cycle, as well as on parasites synchronized in G1 and S phases. As a first step, we curated the current annotation of ribosomal proteins (RPs), assessing copy number, ribosomal location, N- and C-terminal extensions, expression levels, and possible extra-ribosomal roles. Experimentally, Ribo-Seq analysis revealed a global repression of RP mRNA translation in the metacyclic trypomastigote stage. However, some RP transcripts showed distinct behavior, suggesting selective resistance to this repression. Additionally, we applied a multi-omics approach—combining RNA-Seq, Ribo-Seq, and proteomics—on parasites synchronized in G1 and S phases. This analysis uncovered individual differences in translational efficiency of RP mRNAs, as well as in the steady-state protein levels of RPs. These findings support the idea that the composition of the translational machinery may vary during these biological transitions. To further explore this hypothesis, we conducted quantitative proteomic analysis of ribosome-enriched fractions obtained from epimastigotes, metacyclic trypomastigotes, and cell cycle–synchronized parasites in G1 and S phases. Through these experiments, we found that certain ribosomal proteins exhibit differential abundance in ribosome-enriched fractions across the different life cycle stages. We also observed variations in the levels of proteins associated with ribosomes. The differential abundance of components of the translational machinery could have regulatory implications, representing a potential new layer of gene expression regulation in T. cruzi.eng4th Molecular Biosystems Conference: Eukaryotic Gene Regulation & Functional Genomics, Puerto Varas, 09/2025reponame:IIBCE en REDIinstname:Instituto de Investigaciones Biológicas Clemente Estableinstacron:Instituto de Investigaciones Biológicas Clemente EstableTrypanosoma cruziRibosomaRegulación de la expresión génicaCiencias Naturales y ExactasCiencias BiológicasBioquímica y Biología MolecularDifferential composition of the translational machinery of Trypanosoma cruzi during metacyclogenesis and cell cycleDocumento de conferenciaPublicadoinfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/conferenceObject//Ciencias Naturales y Exactas/Ciencias Biológicas/Bioquímica y Biología MolecularRivara, MartínRadío, SantiagoChávez, SantiagoDuhagon, María AnaSmircich, PabloSotelo, JoséLICENSElicense.txtlicense.txttext/plain; charset=utf-85151https://redi.anii.org.uy/jspui/bitstream/20.500.12381/5357/2/license.txtfcc8b2ddb26c820405a37f9e0cae7f18MD52ORIGINALAbstract MolBio 2025 RIVARA.pdfAbstract MolBio 2025 RIVARA.pdfapplication/pdf68629https://redi.anii.org.uy/jspui/bitstream/20.500.12381/5357/1/Abstract%20MolBio%202025%20RIVARA.pdfe57a516e558144ff4ccd80752c8061b2MD5120.500.12381/53572026-01-12 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científico-tecnológicohttps://www.gub.uy/ministerio-educacion-cultura/iibcehttps://redi.anii.org.uy/oai/requestvcarballo@iibce.edu.uyUruguayopendoar:9421_32026-01-12T18:27:06IIBCE en REDI - Instituto de Investigaciones Biológicas Clemente Establefalse |
| spellingShingle | Differential composition of the translational machinery of Trypanosoma cruzi during metacyclogenesis and cell cycle Rivara, Martín Trypanosoma cruzi Ribosoma Regulación de la expresión génica Ciencias Naturales y Exactas Ciencias Biológicas Bioquímica y Biología Molecular |
| status_str | publishedVersion |
| title | Differential composition of the translational machinery of Trypanosoma cruzi during metacyclogenesis and cell cycle |
| title_full | Differential composition of the translational machinery of Trypanosoma cruzi during metacyclogenesis and cell cycle |
| title_fullStr | Differential composition of the translational machinery of Trypanosoma cruzi during metacyclogenesis and cell cycle |
| title_full_unstemmed | Differential composition of the translational machinery of Trypanosoma cruzi during metacyclogenesis and cell cycle |
| title_short | Differential composition of the translational machinery of Trypanosoma cruzi during metacyclogenesis and cell cycle |
| title_sort | Differential composition of the translational machinery of Trypanosoma cruzi during metacyclogenesis and cell cycle |
| topic | Trypanosoma cruzi Ribosoma Regulación de la expresión génica Ciencias Naturales y Exactas Ciencias Biológicas Bioquímica y Biología Molecular |
| url | https://hdl.handle.net/20.500.12381/5357 |