Abstract
The nonsense-mediated mRNA decay (NMD) pathway was initially identified as a surveillance pathway that degrades mRNAs containing premature termination codons (PTCs). NMD is now also recognized as a post-transcriptional regulatory pathway that regulates the expression of natural mRNAs. Earlier studies demonstrated that regulation of functionally related natural mRNAs by NMD can be differential and condition-specific in Saccharomyces cerevisiae. Here, we investigated the regulation of MAC1 mRNAs by NMD in response to copper as well as the role the MAC1 3′-UTR plays in this regulation. MAC1 is a copper-sensing transcription factor that regulates the high-affinity copper uptake system. MAC1 expression is activated upon copper deprivation. We found that MAC1 mRNAs are regulated by NMD under complete minimal (CM) but escaped NMD under low and high copper conditions. Mac1 protein regulated gene, CTR1 is not regulated by NMD in conditions where MAC1 mRNAs are NMD sensitive. We also found that the MAC1 3′-UTR is the NMD targeting feature on the mRNAs, and that MAC1 mRNAs lacking 3′-UTRs were stabilized during copper deprivation. Our results demonstrate a mechanism of regulation for a metal-sensing transcription factor, at both the post-transcriptional and post-translational levels, where MAC1 mRNA levels are regulated by NMD and copper, while the activity of Mac1p is controlled by copper levels.
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Acknowledgements
We are grateful to Jacqueline Carroll, Ethan Blasdel, and Ziwen He for critically reading the manuscript. We also thank the Molecular Biosciences Center (MBC) facility at Baylor University for equipment and supplies.
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This work was supported by the National Institute of General Medical Sciences of the NIH under Award Number R15GM117524. The content is solely the responsibility of the authors and does not necessarily represent the official views of the National Institutes of Health.
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Xinyi Zhang contributed to the design, data acquisition, data analysis, interpretation, drafting, reviewing and final approval of the manuscript. Bessie Kebaara conceived and designed the experiments, contributed to the preparation of the manuscript, approved the final version and acquired funding for the research.
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Zhang, X., Kebaara, B.W. Nonsense-mediated mRNA decay of metal-binding activator MAC1 is dependent on copper levels and 3′-UTR length in Saccharomyces cerevisiae. Curr Genet 70, 5 (2024). https://doi.org/10.1007/s00294-024-01291-9
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DOI: https://doi.org/10.1007/s00294-024-01291-9