Trypanosoma cruzi assembling and centromeres
Trypanosoma cruzi , the etiological agent of Chagas disease, exhibits peculiarities in genome organization and gene expression, such as polycistronic transcription, trans-splicing, and predominantly post-transcriptional regulation . Mitosis in T. cruzi occurs with semi-condensation of chromosomes, which prevents the use of classical cytogenetics to determine the exact number of chromosomes. Among the genomic features of T. cruzi, centromeric regions are poorly understood, despite their importance in chromosome segregation and their impact on genome architecture. Genome organization patterns, including those associated with centromeric regions, can be detected using Chromosome Conformation Capture (3C) techniques, such as Hi-C. Understanding how chromosomes are organized and compacted can provide insights into the complex relationships between nuclear organization and centromere positioning. Located in centromeric regions are kinetochores, which are macromolecular complexes that govern chromosome segregation. None of the canonical kinetochore proteins have been identified in trypanosomatids; however, KKT proteins (Kinetoplastid Kinetochore Proteins), found exclusively in trypanosomatids, have been described as present in kinetochores. In this study, we explored the putative centromeric regions including the generation of tagged (mNeonGreen-Myc) strains for two KKT proteins, KKT2 and KKT3, followed by genomic localization using ChIP-seq, and through the generation of high-coverage Hi-C data which revealed the 3D organization of centromeres.