Fission yeast RNA‐binding proteins Puf2 and Puf4 are involved in repression of ferrireductase Frp1 expression in response to iron
| Autor: | Eric Massé, François Bachand, Darren Henry, Vincent Normant, Simon Labbé, Gordon Chua, Jude Beaudoin, Ariane Brault |
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| Rok vydání: | 2021 |
| Předmět: |
Untranslated region
FMN Reductase Iron RNA-binding protein Biology Microbiology Gene Expression Regulation Fungal Schizosaccharomyces RNA Messenger RNA Processing Post-Transcriptional DNA Fungal Promoter Regions Genetic 3' Untranslated Regions Molecular Biology Psychological repression chemistry.chemical_classification Messenger RNA Reporter gene RNA-Binding Proteins biology.organism_classification Reverse transcriptase Cell biology Enzyme chemistry Mutation Schizosaccharomyces pombe Schizosaccharomyces pombe Proteins |
| Zdroj: | Molecular Microbiology. 116:1361-1377 |
| ISSN: | 1365-2958 0950-382X |
| Popis: | This study identifies a post-transcriptional mechanism of iron uptake regulation by Puf2 and Puf4 of the Pumilio and FBF (Puf) family of RNA-binding proteins in Schizosaccharomyces pombe. Cells expressing Puf2 and Puf4 stimulate decay of the frp1+ mRNA encoding a key enzyme of the reductive iron uptake pathway. Results consistently showed that frp1+ mRNA is stabilized in puf2Δ puf4Δ mutant cells under iron-replete conditions. As a result, puf2Δ puf4Δ cells exhibit an increased sensitivity to iron accompanied by enhanced ferrireductase activity. A pool of GFP-frp1+ 3'UTR RNAs was generated using a reporter gene containing the 3' untranslated region (UTR) of frp1+ that was under the control of a regulatable promoter. Results showed that Puf2 and Puf4 accelerate the destabilization of mRNAs containing the frp1+ 3'UTR which harbors two Pumilio response elements (PREs). Binding studies revealed that the PUM-homology RNA-binding domain of Puf2 and Puf4 expressed in Escherichia coli specifically interacts with PREs in the frp1+ 3'UTR. Using RNA immunoprecipitation in combination with reverse transcription qPCR assays, results showed that Puf2 and Puf4 interact preferentially with frp1+ mRNA under basal and iron-replete conditions, thereby contributing to inhibit Frp1 production and protecting cells against toxic levels of iron. |
| Databáze: | OpenAIRE |
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