Chromatin Proteomics Reveals Variable Histone Modifications during the Life Cycle of Trypanosoma cruzi.
Histones are well-conserved proteins that form the basic structure of chromatin in eukaryotes and undergo several post-translational modifications, which are important for the control of transcription, replication, DNA damage repair, and chromosome condensation. In early branched organisms, histones are less conserved and appear to contain alternative sites for modifications, which could reveal evolutionary unique functions of histone modifications in gene expression and other chromatin-based processes. Here, by using high-resolution mass spectrometry, we identified and quantified histone post-translational modifications in two life cycle stages of Trypanosoma cruzi, the protozoan parasite that causes Chagas disease. We detected 44 new modifications, namely: 18 acetylations, seven monomethylations, seven dimethylations, seven trimethylations, and four phosphorylations. We found that replicative (epimastigote stage) contains more histone modifications than nonreplicative and infective parasites (trypomastigote stage). Acetylations of lysines at the C-terminus of histone H2A and methylations of lysine 23 of histone H3 were found to be enriched in trypomastigotes. In contrast, phosphorylation in serine 23 of H2B and methylations of lysine 76 of histone H3 predominates in proliferative states. The presence of one or two methylations in the lysine 76 was found in cells undergoing mitosis and cytokinesis, typical of proliferating parasites. Our findings provide new insights into the role of histone modifications related to the control of gene expression and cell-cycle regulation in an early divergent organism.
Laboratório Especial de Ciclo Celular, Center of Toxins, Immune Response and Cell Signaling - CeTICS, Instituto Butantan , São Paulo 05503-900, Brazil.|Departamento de Física e Informática, Instituto de Física de São Carlos, Universidade de São Paulo - USP , São Carlos, São Paulo 13563-120, Brazil.
Teresa Cristina Leandro
TC
de Jesus
Departamento de Microbiologia, Imunologia e Parasitologia, Universidade Federal de São Paulo , São Paulo 04039-032, Brazil.
Vinícius Santana
VS
Nunes
Laboratório Especial de Ciclo Celular, Center of Toxins, Immune Response and Cell Signaling - CeTICS, Instituto Butantan , São Paulo 05503-900, Brazil.
Mariana de Camargo
Mde C
Lopes
Departamento de Física e Informática, Instituto de Física de São Carlos, Universidade de São Paulo - USP , São Carlos, São Paulo 13563-120, Brazil.
Daiana Evelin
DE
Martil
Laboratório Especial de Ciclo Celular, Center of Toxins, Immune Response and Cell Signaling - CeTICS, Instituto Butantan , São Paulo 05503-900, Brazil.
Leo Kei
LK
Iwai
Departamento de Microbiologia, Imunologia e Parasitologia, Universidade Federal de São Paulo , São Paulo 04039-032, Brazil.
Nilmar Silvio
NS
Moretti
Departamento de Microbiologia, Imunologia e Parasitologia, Universidade Federal de São Paulo , São Paulo 04039-032, Brazil.
Fabrício Castro
FC
Machado
Departamento de Microbiologia, Imunologia e Parasitologia, Universidade Federal de São Paulo , São Paulo 04039-032, Brazil.
Mariana L
ML
de Lima-Stein
Departamento de Física e Informática, Instituto de Física de São Carlos, Universidade de São Paulo - USP , São Carlos, São Paulo 13563-120, Brazil.
Otavio Henrique
OH
Thiemann
Laboratório Especial de Ciclo Celular, Center of Toxins, Immune Response and Cell Signaling - CeTICS, Instituto Butantan , São Paulo 05503-900, Brazil.
Maria Carolina
MC
Elias
Department of Cell and Developmental Biology, Theodor-Boveri-Institute at the Biocenter, University of Würzburg , 97070 Germany.
Christian
C
Janzen
Departamento de Microbiologia, Imunologia e Parasitologia, Universidade Federal de São Paulo , São Paulo 04039-032, Brazil.
Sergio
S
Schenkman
Laboratório Especial de Ciclo Celular, Center of Toxins, Immune Response and Cell Signaling - CeTICS, Instituto Butantan , São Paulo 05503-900, Brazil.
Julia Pinheiro Chagas
JP
da Cunha
D000107:Acetylation; D002453:Cell Cycle; D002843:Chromatin / Q000737:chemistry*; D005786:Gene Expression Regulation; D042421:Histone Code; D008018:Life Cycle Stages; D008745:Methylation; D010766:Phosphorylation; D011499:Protein Processing, Post-Translational / Q000502:physiology; D040901:Proteomics / Q000379:methods*; D014349:Trypanosoma cruzi
D002843:Chromatin
Trypanosoma cruzi; acetylation; cell cycle; chromatin; histone; life cycle; methylation