Characterising ISWI chromatin remodeler in Trypanosoma cruzi
BACKGROUND Imitation SWItch (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 Trypanosoma cruzi, a protozoan that controls gene expression princ...
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| Format: | Article |
| Language: | English |
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Fundação Oswaldo Cruz (FIOCRUZ)
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| Series: | Memorias do Instituto Oswaldo Cruz |
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| Online Access: | http://www.scielo.br/scielo.php?script=sci_arttext&pid=S0074-02762020000100318&lng=en&tlng=en |
| _version_ | 1797718248017362944 |
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| author | Yirys Díaz-Olmos Michel Batista Adriana Ludwig Fabricio K Marchini |
| author_facet | Yirys Díaz-Olmos Michel Batista Adriana Ludwig Fabricio K Marchini |
| author_sort | Yirys Díaz-Olmos |
| collection | DOAJ |
| description | BACKGROUND Imitation SWItch (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 Trypanosoma cruzi, a protozoan that controls gene expression principally post-transcriptionally, the transcriptional regulation mechanisms mediated by chromatin remodeling are poorly understood. OBJECTIVE To characterise the ISWI remodeler in T. cruzi (TcISWI). METHODS A new version of pTcGW vectors was constructed to express green fluorescent protein (GFP)-tagged TcISWI. CRISPR-Cas9 system was used to obtain parasites with inactivated TcISWI gene and we determined TcISWI partners by cryomilling-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 [nucleoplasmin-like protein (NLP), regulator of chromosome condensation 1-like protein (RCCP) and phenylalanine/tyrosine-rich protein (FYRP)], previously characterised in T. brucei, and new components in TcISWI complex [DRBD2, DHH1 and proteins containing a domain characteristic of structural maintenance of chromosomes (SMC) proteins]. Data are available via ProteomeXchange with identifier PXD017869. 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 TcISWI chromatin remodeler in different nuclear processes in T. cruzi, including mRNA nuclear export control and chromatin compaction. Further work is necessary to clarify the TcISWI functional diversity that arises from this protein interaction study. |
| first_indexed | 2024-03-12T08:48:08Z |
| format | Article |
| id | doaj.art-c8f7603dabd84f2cb4cd360026ab7cb0 |
| institution | Directory Open Access Journal |
| issn | 0074-0276 1678-8060 |
| language | English |
| last_indexed | 2024-03-12T08:48:08Z |
| publisher | Fundação Oswaldo Cruz (FIOCRUZ) |
| record_format | Article |
| series | Memorias do Instituto Oswaldo Cruz |
| spelling | doaj.art-c8f7603dabd84f2cb4cd360026ab7cb02023-09-02T16:25:05ZengFundação Oswaldo Cruz (FIOCRUZ)Memorias do Instituto Oswaldo Cruz0074-02761678-806011510.1590/0074-02760190457S0074-02762020000100318Characterising ISWI chromatin remodeler in Trypanosoma cruziYirys Díaz-OlmosMichel BatistaAdriana LudwigFabricio K MarchiniBACKGROUND Imitation SWItch (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 Trypanosoma cruzi, a protozoan that controls gene expression principally post-transcriptionally, the transcriptional regulation mechanisms mediated by chromatin remodeling are poorly understood. OBJECTIVE To characterise the ISWI remodeler in T. cruzi (TcISWI). METHODS A new version of pTcGW vectors was constructed to express green fluorescent protein (GFP)-tagged TcISWI. CRISPR-Cas9 system was used to obtain parasites with inactivated TcISWI gene and we determined TcISWI partners by cryomilling-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 [nucleoplasmin-like protein (NLP), regulator of chromosome condensation 1-like protein (RCCP) and phenylalanine/tyrosine-rich protein (FYRP)], previously characterised in T. brucei, and new components in TcISWI complex [DRBD2, DHH1 and proteins containing a domain characteristic of structural maintenance of chromosomes (SMC) proteins]. Data are available via ProteomeXchange with identifier PXD017869. 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 TcISWI chromatin remodeler in different nuclear processes in T. cruzi, including mRNA nuclear export control and chromatin compaction. Further work is necessary to clarify the TcISWI functional diversity that arises from this protein interaction study.http://www.scielo.br/scielo.php?script=sci_arttext&pid=S0074-02762020000100318&lng=en&tlng=eniswichromatin remodelinginteracting partnersreverse geneticsptcgw plasmid vectors |
| spellingShingle | Yirys Díaz-Olmos Michel Batista Adriana Ludwig Fabricio K Marchini Characterising ISWI chromatin remodeler in Trypanosoma cruzi Memorias do Instituto Oswaldo Cruz iswi chromatin remodeling interacting partners reverse genetics ptcgw plasmid vectors |
| title | Characterising ISWI chromatin remodeler in Trypanosoma cruzi |
| title_full | Characterising ISWI chromatin remodeler in Trypanosoma cruzi |
| title_fullStr | Characterising ISWI chromatin remodeler in Trypanosoma cruzi |
| title_full_unstemmed | Characterising ISWI chromatin remodeler in Trypanosoma cruzi |
| title_short | Characterising ISWI chromatin remodeler in Trypanosoma cruzi |
| title_sort | characterising iswi chromatin remodeler in trypanosoma cruzi |
| topic | iswi chromatin remodeling interacting partners reverse genetics ptcgw plasmid vectors |
| url | http://www.scielo.br/scielo.php?script=sci_arttext&pid=S0074-02762020000100318&lng=en&tlng=en |
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