Differential composition of the translational machinery of Trypanosoma cruzi during metacyclogenesis and cell cycle

Rivara, Martín - Radío, Santiago - Chávez, Santiago - Duhagon, María Ana - Smircich, Pablo - Sotelo, José

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.

Detalles Bibliográficos
2025
Trypanosoma cruzi
Ribosoma
Regulación de la expresión génica
Ciencias Naturales y Exactas
Ciencias Biológicas
Bioquímica y Biología Molecular
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