Improving genome-wide mapping of nucleosomes in Trypanosome cruzi.

BEATI, PAULA; MASSIMINO STEPÑICKA, MILENA; VILCHEZ LARREA, SALOMÉ C.; SMIRCICH, PABLO; ALONSO, GUILLERMO D.; OCAMPO, JOSEFINA

PLOS ONE

PUBLIC LIBRARY SCIENCE

Año: 2023 vol. 18

1932-6203

In Trypanosoma cruziDNA is packaged into chromatin by octamers of histone proteins that form nucleosomes. Transcription of protein coding genes in trypanosomes is constitutive producing polycistronic units and gene expression is primarily regulated post-transcription- ally. However, chromatin organization influences DNA dependent processes. Hence, deter- mining nucleosome position is of uppermost importance to understand the peculiarities found in trypanosomes. To map nucleosomes genome-wide in several organisms, digestion of chromatin with micrococcal nuclease followed by deep sequencing has been applied. Nonetheless, the special requirements for cell manipulation and the uniqueness of the chro- matin organization in trypanosomes entails a customized analytical approach. In this work, we adjusted this broadly used method to the hybrid reference strain, CL Brener. Particularly, we implemented an exhaustive and thorough computational workflow to overcome the diffi- culties imposed by this complex genome. We tested the performance of two aligners, Bow tie2 and HISAT2, and discuss their advantages and caveats. Specifically, we highlight the relevance of using the whole genome as a reference instead of the commonly used Esmer- aldo-like haplotype to avoid spurious alignments. Additionally, we show that using the whole genome refines the average nucleosome representation, but also the quality of mapping for every region represented. Moreover, we show that the average nucleosome organization around trans-splicing acceptor site described before, is not just an average since the same chromatin pattern is detected for most of the represented regions. In addition, we extended the study to a non-hybrid strain applying the experimental and analytical approach to Sylvio- X10 strain. Furthermore, we provide a source code for the construction of 2D plots and heat- maps which are easy to adapt to any T. cruzi strain.