Mol Microbiol. 2016 Aug 20. doi: 10.1111/mmi.13485. [Epub ahead of print]
Serpeloni M1,2,3,
Jiménez-Ruiz E3,
Medeiros Vidal N4,
Kroeber C3,
Andenmatten N3,
Lemgruber L3,
Mörking P1,
Pall GS3,
Meissner M3,
Ávila AR1.
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. This article is protected by copyright. All rights reserved.
© 2016 John Wiley & Sons Ltd.
KEYWORDS:
Key-words: mRNA export pathway; Toxoplasma gondii; conditional Cas9 system
- [PubMed - as supplied by publisher]
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