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| Status |
Public on Jun 17, 2020 |
| Title |
Global changes in nitration levels and DNA binding profile of Trypanosoma cruzi histones induced by incubation with host extracellular matrix |
| Organism |
Trypanosoma cruzi |
| Experiment type |
Genome binding/occupancy profiling by high throughput sequencing
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| Summary |
Adhesion of T. cruzi trypomastigotes to components of the extracellular matrix (ECM) is an important step in mammalian host cell invasion. We have recently described a significant increase in the tyrosine nitration levels of histones H2A and H4 when trypomastigotes are incubated with components of the ECM. In this work, we used chromatin immunoprecipitation (ChIP) with an anti-nitro-tyrosine antibody followed by mass spectrometry to identify nitrated DNA binding proteins in T. cruzi and to detect alterations in nitration levels induced upon parasite incubation with the ECM. Histone H1, H2B, H2A and H3 were detected among the 9 most abundant nitrated DNA binding proteins using this proteomic approach. One nitrated tyrosine residue (Y29) was identified in Histone H2B in the MS/MS spectrum. In addition, we observed a significant increase in the nitration levels of histones H1, H2B, H2A and H4 upon parasite incubation with ECM. Finally, we used ChIP-Seq to map global changes in the DNA binding profile of nitrated proteins. We observed a significant change in the binding pattern of nitrated proteins to DNA after parasite incubation with ECM. This work provides the first global profile of nitrated DNA binding proteins in T. cruzi and additional evidence for modification in the nitration profile of histones upon parasite incubation with ECM. Our data also indicate that the interaction of the parasite with the ECM induces alterations in chromatin structure, possibly affecting nuclear functions.
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| Overall design |
Examination of the changes of nitrated proteins interactions with chromatin in T.cruzi after incubation with host extracelular matrix (ECM). Two samples of the parasite incubated or not with ECM (TyM and Ty) were prepared in replicates (Rep1 and Rep2) to be submitted to ChIP-Seq technique. Paired-end experiments generate files R1 and R2.
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| Contributor(s) |
Magalhães RD, Mattos EC, Rozanski A, Galante PA, Palmisano G, Cruz AK, Colli W, Camargo AA, Alves MJ |
| Citation(s) |
32469928 |
| |
| Submission date |
Nov 12, 2019 |
| Last update date |
Jun 17, 2020 |
| Contact name |
Rubens Daniel Miserani Magalhães |
| E-mail(s) |
criphg@gmail.com
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| Organization name |
Ribeirão Preto Medical School
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| Department |
Department of Cellular and Molecular Biology and Pathogenic Bioagents
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| Lab |
Cellular and Molecular Parasitology
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| Street address |
Av. Bandeirantes 3900
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| City |
Ribeirão Preto |
| State/province |
SP |
| ZIP/Postal code |
14049-900 |
| Country |
Brazil |
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| Platforms (1) |
| GPL27740 |
Illumina NextSeq 500 (Trypanosoma cruzi) |
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| Samples (4)
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| Relations |
| BioProject |
PRJNA589041 |
| SRA |
SRP229717 |
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