Unveiling the partners of the DRBD2-mRNP complex, an RBP in Trypanosoma cruzi and ortholog to the yeast SR-protein Gbp2
Autor: | Helisa H. Wippel, Juliane S. Malgarin, Lysangela R. Alves, Paulo C. Carvalho, Alexandre Haruo Inoue, Samuel Goldenberg, Felipe da Veiga Leprevost |
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Jazyk: | angličtina |
Rok vydání: | 2019 |
Předmět: |
Proteomics
Microbiology (medical) Saccharomyces cerevisiae Proteins Immunoprecipitation GBP2 RNA-binding protein Trypanosoma cruzi Protozoan Proteins lcsh:QR1-502 Biology Microbiology lcsh:Microbiology 03 medical and health sciences SR protein Protein Domains Shotgun proteomics Regulation of gene expression Gene expression regulation RNA metabolism 0303 health sciences Messenger RNA Sequence Homology Amino Acid Sequence Analysis RNA 030306 microbiology RNA-Binding Proteins Cell biology Ribonucleoproteins Biosynthetic process Protein Binding Research Article |
Zdroj: | BMC Microbiology, Vol 19, Iss 1, Pp 1-12 (2019) BMC Microbiology |
ISSN: | 1471-2180 |
DOI: | 10.1186/s12866-019-1505-8 |
Popis: | Background RNA-binding proteins (RBPs) are well known as key factors in gene expression regulation in eukaryotes. These proteins associate with mRNAs and other proteins to form mRNP complexes that ultimately determine the fate of target transcripts in the cell. This association is usually mediated by an RNA-recognition motif (RRM). In the case of trypanosomatids, these proteins play a paramount role, as gene expression regulation is mostly posttranscriptional. Despite their relevance in the life cycle of Trypanosoma cruzi, the causative agent of Chagas’ disease, to date, few RBPs have been characterized in this parasite. Results We investigated the role of DRBD2 in T. cruzi, an RBP with two RRM domains that is associated with cytoplasmic translational complexes. We show that DRBD2 is an ortholog of the Gbp2 in yeast, an SR-rich protein involved in mRNA quality control and export. We used an immunoprecipitation assay followed by shotgun proteomics and RNA-seq to assess the interaction partners of the DRBD2-mRNP complex in epimastigotes. The analysis identified mostly proteins involved in RNA metabolism and regulation, such as ALBA1, ALBA3, ALBA4, UBP1, UBP2, DRBD3, and PABP2. The RNA-seq results showed that most of the transcripts regulated by the DRBD2 complex mapped to hypothetical proteins related to multiple processes, such as to biosynthetic process, DNA metabolic process, protein modification, and response to stress. Conclusions The identification of regulatory proteins in the DRBD2-mRNP complex corroborates the important role of DRBD2 in gene expression regulation in T. cruzi. We consider these results an important contribution to future studies regarding gene expression regulation in T. cruzi, especially in the field of RNA-binding proteins. Electronic supplementary material The online version of this article (10.1186/s12866-019-1505-8) contains supplementary material, which is available to authorized users. |
Databáze: | OpenAIRE |
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