Metacyclogenesis defects and gene expression hallmarks of histone deacetylase 4-deficient Trypanosoma cruzi cells

Cassiano Martin Batista,Arthur Schveitzer Ferreira,Gisele Fernanda Assine Picchi-Constante,Michel Batista,Sergio Verjovski-Almeida,Murilo Sena Amaral,Samuel Goldenberg,Eloise Pavão Guerra-Slompo,Nilson Ivo Tonin Zanchin,Ana Carolina Tahira,Monica Visnieski Alcantara

doi:10.1038/s41598-021-01080-1

Cassiano Martin Batista, Arthur Schveitzer Ferreira + Show 9 more

Open Access

https://doi.org/10.1038/s41598-021-01080-1

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Journal: Scientific Reports	Publication Date: Nov 4, 2021
Citations: 3	License type: open-access

Affiliation: Fundação Carlos Chagas, Instituto Butantan

Abstract

Trypanosoma cruzi—the causative agent of Chagas disease—like other kinetoplastids, relies mostly on post-transcriptional mechanisms for regulation of gene expression. However, trypanosomatids undergo drastic changes in nuclear architecture and chromatin structure along their complex life cycle which, combined with a remarkable set of reversible histone post-translational modifications, indicate that chromatin is also a target for control of gene expression and differentiation signals in these organisms. Chromatin-modifying enzymes have a direct impact on gene expression programs and DNA metabolism. In this work, we have investigated the function of T. cruzi histone deacetylase 4 (TcHDAC4). We show that, although TcHDAC4 is not essential for viability, metacyclic trypomastigote TcHDAC4 null mutants show a thin cell body and a round and less condensed nucleus located very close to the kinetoplast. Sixty-four acetylation sites were quantitatively evaluated, which revealed H2AT85ac, H4K10ac and H4K78ac as potential target sites of TcHDAC4. Gene expression analyses identified three chromosomes with overrepresented regions of differentially expressed genes in the TcHDAC4 knockout mutant compared with the wild type, showing clusters of either up or downregulated genes. The adjacent chromosomal location of some of these genes indicates that TcHDAC4 participates in gene expression regulation during T. cruzi differentiation.

Highlights

Trypanosoma cruzi—the causative agent of Chagas disease—like other kinetoplastids, relies mostly on post-transcriptional mechanisms for regulation of gene expression
T. cruzi histone deacetylase 4 (TcHDAC4) is expressed in all four stages tested (Fig. 1A) with higher expression levels occurring in the ones that represent the insect stages, while lower levels are observed in the stages that occur within the mammalian host (Fig. 1A)
We showed that the TcHDAC4 is expressed throughout the T. cruzi life cycle, at higher levels in the developmental stages occurring in the invertebrate vector

Summary

Introduction

Trypanosoma cruzi—the causative agent of Chagas disease—like other kinetoplastids, relies mostly on post-transcriptional mechanisms for regulation of gene expression. Trypanosomatids undergo drastic changes in nuclear architecture and chromatin structure along their complex life cycle which, combined with a remarkable set of reversible histone post-translational modifications, indicate that chromatin is a target for control of gene expression and differentiation signals in these organisms. Chromatin-modifying enzymes are major players in epigenetic mechanisms, mediating reversible covalent modifications and having a direct impact on gene expression programs[3,4]. All T. cruzi canonical and variant histones are decorated with a multitude of post-translational modifications (PTMs) both at the flexible tails and in the globular region[6,7,8,9,10,11] Some of these PTMs correlate with those described for other organisms, while others seem to be exclusive of trypanosomatids, raising interesting questions about their role in epigenetic mechanisms. In the present work, we focus on the characterization of T. cruzi histone deacetylase 4 (TcHDAC4)

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