Mem Inst Oswaldo Cruz, Rio de Janeiro, VOLUME 115 | MARCH 2020
Characterizing ISWI chromatin remodelerin Trypanosomacruzi [ACCEPTED ARTICLES / PRELIMINARY VERSION]
1Laboratório de Ciências e Tecnologias Aplicadas em Saúde, Instituto Carlos Chagas, Fiocruz, Curitiba, PR, Brasil
2Fundación Universidad del Norte, División Ciencias de la Salud, Barranquilla, Colombia
3Mass Spectrometry Facility - RPT02H, Instituto Carlos Chagas, Fiocruz, Curitiba, PR, Brasil.
BACKGROUND: ISWI ATPase is the catalytic subunit in diverse chromatin remodeling complexes. These complexes modify histone-DNA interactions and therefore play a pivotal role in different DNA-dependent processes. In Trypanosomacruzi, a protozoan that controls gene expression principally post-transcriptionally, the transcriptional regulation mechanisms mediated by chromatin remodeling are poorly understood.
OBJECTIVE: Tocharacterize the ISWI remodeler in T.cruzi(TcISWI).
METHODS: A new version of pTcGW vectors was constructed to express GFP-tagged TcISWI. CRISPR-Cas9 system was used to obtain parasites with inactivated TcISWI gene and we determined TcISWI partners bycryomilling-affinity purification-mass spectrometry (MS) assay as an approximation to start to unravel the function of this protein.
FINDINGS: Our approach identified known ISWI partners (NLP, RCCP and FYRP), previously characterized in T. brucei, and new components in TcISWI complex (DRBD2, DHH1 and proteins containing SMC domains).
MAIN CONCLUSIONS: In addition to its participation in transcriptional silencing, as it was reported in T. brucei, the data generated here provide a framework that suggests a role for TcISWIchromatin remodeler in different nuclear processes in T. cruzi, including mRNA nuclear export control and chromatin compaction. Further work isnecessary to clarify the TcISWI functional diversity that arises from this protein interaction study.