4.5 Article

UAP56 is a conserved crucial component of a divergent mRNA export pathway in Toxoplasma gondii

Journal

MOLECULAR MICROBIOLOGY
Volume 102, Issue 4, Pages 672-689

Publisher

WILEY
DOI: 10.1111/mmi.13485

Keywords

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Funding

  1. Programa Estrategico de Apoio a Pesquisa em Saude (PAPES/FIOCRUZ, Brazil) [407775/2012-9 - CNPq]
  2. Fundacao Araucaria [145/2015]
  3. CAPES Foundation, Ministry of Education of Brazil (CAPES, Brazil) [PDSE Fellowship] [9445-12-9]
  4. National Counsel of Technological and Scientific Development (CNPq, Brazil) [PDE Fellowship] [249741/2013-0]
  5. ERC-Starting grant [ERC-2012-StG309255-EndoTox]
  6. Wellcome Trust [087582/Z/08/Z, 085349]
  7. Intramural Research Program of the National Library of Medicine, National Institutes of Health
  8. Conselho Nacional de Desenvolvimento Cientifico e Tecnologico
  9. National Institutes of Health

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Nucleo-cytoplasmic RNA export is an essential post-transcriptional step to control gene expression in eukaryotic cells and is poorly understood in apicomplexan parasites. With the exception of UAP56, a component of TREX (Transcription Export) complex, other components of mRNA export machinery are not well conserved in divergent supergroups. Here, we use Toxoplasma gondii as a model system to functionally characterize TgUAP56 and its potential interaction factors. We demonstrate that TgUAP56 is crucial for mRNA export and that functional interference leads to significant accumulation of mRNA in the nucleus. It was necessary to employ bioinformatics and phylogenetic analysis to identify orthologs related to mRNA export, which show a remarkable low level of conservation in T. gondii. We adapted a conditional Cas9/CRISPR system to carry out a genetic screen to verify if these factors were involved in mRNA export in T. gondii. Only the disruption of TgRRM_1330 caused accumulation of mRNA in the nucleus as found with TgUAP56. This protein is potentially a divergent partner of TgUAP56, and provides insight into a divergent mRNA export pathway in apicomplexans.

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