Stage-specific MCM protein expression in Trypanosoma cruzi: insights into metacyclogenesis and G1 arrested epimastigotes

Trypanosoma cruzi is a protozoan parasite that is the etiological agent of Chagas disease, which is endemic to Latin America with reported cases in non-endemic regions such as Europe, Asia, and Oceania due to migration. During its lifecycle, T. cruzi alternates between replicative and non-replicativ...

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Main Authors: Bruno Alves Santarossa, Évelin Mariani, Artur da Paixão Corrêa, Fernanda C. Costa, Martin C. Taylor, John M. Kelly, Maria Carolina Elias, Simone Guedes Calderano
Format: Article
Language:English
Published: Frontiers Media S.A. 2025-05-01
Series:Frontiers in Cellular and Infection Microbiology
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Online Access:https://www.frontiersin.org/articles/10.3389/fcimb.2025.1584812/full
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author Bruno Alves Santarossa
Évelin Mariani
Artur da Paixão Corrêa
Fernanda C. Costa
Martin C. Taylor
John M. Kelly
Maria Carolina Elias
Simone Guedes Calderano
author_facet Bruno Alves Santarossa
Évelin Mariani
Artur da Paixão Corrêa
Fernanda C. Costa
Martin C. Taylor
John M. Kelly
Maria Carolina Elias
Simone Guedes Calderano
author_sort Bruno Alves Santarossa
collection DOAJ
description Trypanosoma cruzi is a protozoan parasite that is the etiological agent of Chagas disease, which is endemic to Latin America with reported cases in non-endemic regions such as Europe, Asia, and Oceania due to migration. During its lifecycle, T. cruzi alternates between replicative and non-replicative infective lifeforms. Metacyclogenesis is the most studied transition of the T. cruzi life cycle, where replicative epimastigotes differentiate into infective metacyclic trypomastigotes inside the gut of the triatomine vector. This early-branching organism expresses a divergent pre-replication complex (pre-RC) where the only conserved component is the MCM2–7 protein family. Given the role of pre-RC components in cell cycle regulation, we investigated whether MCM expression and location could be involved in proliferation control in epimastigotes and during metacyclogenesis. Using CRISPR/Cas9, we tagged MCM subunits and tracked their expression and subcellular localization. Our findings reveal that MCM subunits are consistently expressed and localized to the nucleus throughout the epimastigote cell cycle, including in G1/G0-arrested cells. However, MCM subunits are degraded during metacyclogenesis as cells enter the G0 state, marking the transition to replication arrest. Therefore, epimastigotes arrested in G1/G0 can either maintain MCM complex expression and resume the cell cycle when conditions become favorable, or they can undergo metacyclogenesis, exiting the cell cycle and entering a G0 state, where MCM subunits are degraded as part of the replication repression mechanism.
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spelling doaj-art-ceacdb687f10458da6570c5331c149f12025-08-20T03:48:23ZengFrontiers Media S.A.Frontiers in Cellular and Infection Microbiology2235-29882025-05-011510.3389/fcimb.2025.15848121584812Stage-specific MCM protein expression in Trypanosoma cruzi: insights into metacyclogenesis and G1 arrested epimastigotesBruno Alves Santarossa0Évelin Mariani1Artur da Paixão Corrêa2Fernanda C. Costa3Martin C. Taylor4John M. Kelly5Maria Carolina Elias6Simone Guedes Calderano7Cell Cycle Laboratory, Butantan Institute, São Paulo, BrazilCell Cycle Laboratory, Butantan Institute, São Paulo, BrazilCell Cycle Laboratory, Butantan Institute, São Paulo, BrazilDepartment of Infection Biology, London School of Hygiene and Tropical Medicine, London, United KingdomDepartment of Infection Biology, London School of Hygiene and Tropical Medicine, London, United KingdomDepartment of Infection Biology, London School of Hygiene and Tropical Medicine, London, United KingdomCell Cycle Laboratory, Butantan Institute, São Paulo, BrazilCell Cycle Laboratory, Butantan Institute, São Paulo, BrazilTrypanosoma cruzi is a protozoan parasite that is the etiological agent of Chagas disease, which is endemic to Latin America with reported cases in non-endemic regions such as Europe, Asia, and Oceania due to migration. During its lifecycle, T. cruzi alternates between replicative and non-replicative infective lifeforms. Metacyclogenesis is the most studied transition of the T. cruzi life cycle, where replicative epimastigotes differentiate into infective metacyclic trypomastigotes inside the gut of the triatomine vector. This early-branching organism expresses a divergent pre-replication complex (pre-RC) where the only conserved component is the MCM2–7 protein family. Given the role of pre-RC components in cell cycle regulation, we investigated whether MCM expression and location could be involved in proliferation control in epimastigotes and during metacyclogenesis. Using CRISPR/Cas9, we tagged MCM subunits and tracked their expression and subcellular localization. Our findings reveal that MCM subunits are consistently expressed and localized to the nucleus throughout the epimastigote cell cycle, including in G1/G0-arrested cells. However, MCM subunits are degraded during metacyclogenesis as cells enter the G0 state, marking the transition to replication arrest. Therefore, epimastigotes arrested in G1/G0 can either maintain MCM complex expression and resume the cell cycle when conditions become favorable, or they can undergo metacyclogenesis, exiting the cell cycle and entering a G0 state, where MCM subunits are degraded as part of the replication repression mechanism.https://www.frontiersin.org/articles/10.3389/fcimb.2025.1584812/fullMini-Chromosome MaintenanceMCMTrypanosoma cruzimetacyclogenesisreplication controlG0
spellingShingle Bruno Alves Santarossa
Évelin Mariani
Artur da Paixão Corrêa
Fernanda C. Costa
Martin C. Taylor
John M. Kelly
Maria Carolina Elias
Simone Guedes Calderano
Stage-specific MCM protein expression in Trypanosoma cruzi: insights into metacyclogenesis and G1 arrested epimastigotes
Frontiers in Cellular and Infection Microbiology
Mini-Chromosome Maintenance
MCM
Trypanosoma cruzi
metacyclogenesis
replication control
G0
title Stage-specific MCM protein expression in Trypanosoma cruzi: insights into metacyclogenesis and G1 arrested epimastigotes
title_full Stage-specific MCM protein expression in Trypanosoma cruzi: insights into metacyclogenesis and G1 arrested epimastigotes
title_fullStr Stage-specific MCM protein expression in Trypanosoma cruzi: insights into metacyclogenesis and G1 arrested epimastigotes
title_full_unstemmed Stage-specific MCM protein expression in Trypanosoma cruzi: insights into metacyclogenesis and G1 arrested epimastigotes
title_short Stage-specific MCM protein expression in Trypanosoma cruzi: insights into metacyclogenesis and G1 arrested epimastigotes
title_sort stage specific mcm protein expression in trypanosoma cruzi insights into metacyclogenesis and g1 arrested epimastigotes
topic Mini-Chromosome Maintenance
MCM
Trypanosoma cruzi
metacyclogenesis
replication control
G0
url https://www.frontiersin.org/articles/10.3389/fcimb.2025.1584812/full
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