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Characterization of the transactivation and nuclear localization functions of Pichia pastoris zinc finger transcription factor Mxr1p
The zinc finger transcription factor Mxr1p regulates the transcription of genes involved in methanol, acetate, and amino acid metabolism of the industrial yeast Pichia pastoris (a.k.a. Komagataella phaffii) by binding to Mxr1p response elements in their promoters. Here, we demonstrate that Mxr1p is...
Autores principales: | , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
American Society for Biochemistry and Molecular Biology
2021
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8526985/ https://www.ncbi.nlm.nih.gov/pubmed/34582889 http://dx.doi.org/10.1016/j.jbc.2021.101247 |
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author | Gupta, Aditi Krishna Rao, Kamisetty Sahu, Umakant Rangarajan, Pundi N. |
author_facet | Gupta, Aditi Krishna Rao, Kamisetty Sahu, Umakant Rangarajan, Pundi N. |
author_sort | Gupta, Aditi |
collection | PubMed |
description | The zinc finger transcription factor Mxr1p regulates the transcription of genes involved in methanol, acetate, and amino acid metabolism of the industrial yeast Pichia pastoris (a.k.a. Komagataella phaffii) by binding to Mxr1p response elements in their promoters. Here, we demonstrate that Mxr1p is a key regulator of ethanol metabolism as well. Using transcriptomic analysis, we identified target genes of Mxr1p that mediate ethanol metabolism, including ALD6-1 encoding an aldehyde dehydrogenase. ALD6-1 is essential for ethanol metabolism, and the ALD6-1 promoter harbors three Mxr1p response elements to which Mxr1p binds in vitro and activates transcription in vivo. We show that a nine-amino acid transactivation domain located between amino acids 365 and 373 of Mxr1p is essential for the transactivation of ALD6-1 to facilitate ethanol metabolism. Mxr1N250, containing the N-terminal 250 amino acids of Mxr1p, localized to the nucleus of cells metabolizing ethanol dependent on basic amino acid residues present between amino acids 75 and 85. While the N-terminal 400 amino acids of Mxr1p are sufficient for the activation of target genes essential for ethanol metabolism, the region between amino acids 401 and 1155 was also required for the regulation of genes essential for methanol metabolism. Finally, we identified several novel genes whose expression is differentially regulated by Mxr1p during methanol metabolism by DNA microarray. This study demonstrates that Mxr1p is a key regulator of ethanol metabolism and provides new insights into the mechanism by which Mxr1p functions as a global regulator of multiple metabolic pathways of P. pastoris. |
format | Online Article Text |
id | pubmed-8526985 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2021 |
publisher | American Society for Biochemistry and Molecular Biology |
record_format | MEDLINE/PubMed |
spelling | pubmed-85269852021-10-25 Characterization of the transactivation and nuclear localization functions of Pichia pastoris zinc finger transcription factor Mxr1p Gupta, Aditi Krishna Rao, Kamisetty Sahu, Umakant Rangarajan, Pundi N. J Biol Chem Research Article The zinc finger transcription factor Mxr1p regulates the transcription of genes involved in methanol, acetate, and amino acid metabolism of the industrial yeast Pichia pastoris (a.k.a. Komagataella phaffii) by binding to Mxr1p response elements in their promoters. Here, we demonstrate that Mxr1p is a key regulator of ethanol metabolism as well. Using transcriptomic analysis, we identified target genes of Mxr1p that mediate ethanol metabolism, including ALD6-1 encoding an aldehyde dehydrogenase. ALD6-1 is essential for ethanol metabolism, and the ALD6-1 promoter harbors three Mxr1p response elements to which Mxr1p binds in vitro and activates transcription in vivo. We show that a nine-amino acid transactivation domain located between amino acids 365 and 373 of Mxr1p is essential for the transactivation of ALD6-1 to facilitate ethanol metabolism. Mxr1N250, containing the N-terminal 250 amino acids of Mxr1p, localized to the nucleus of cells metabolizing ethanol dependent on basic amino acid residues present between amino acids 75 and 85. While the N-terminal 400 amino acids of Mxr1p are sufficient for the activation of target genes essential for ethanol metabolism, the region between amino acids 401 and 1155 was also required for the regulation of genes essential for methanol metabolism. Finally, we identified several novel genes whose expression is differentially regulated by Mxr1p during methanol metabolism by DNA microarray. This study demonstrates that Mxr1p is a key regulator of ethanol metabolism and provides new insights into the mechanism by which Mxr1p functions as a global regulator of multiple metabolic pathways of P. pastoris. American Society for Biochemistry and Molecular Biology 2021-09-25 /pmc/articles/PMC8526985/ /pubmed/34582889 http://dx.doi.org/10.1016/j.jbc.2021.101247 Text en © 2021 The Authors https://creativecommons.org/licenses/by/4.0/This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/). |
spellingShingle | Research Article Gupta, Aditi Krishna Rao, Kamisetty Sahu, Umakant Rangarajan, Pundi N. Characterization of the transactivation and nuclear localization functions of Pichia pastoris zinc finger transcription factor Mxr1p |
title | Characterization of the transactivation and nuclear localization functions of Pichia pastoris zinc finger transcription factor Mxr1p |
title_full | Characterization of the transactivation and nuclear localization functions of Pichia pastoris zinc finger transcription factor Mxr1p |
title_fullStr | Characterization of the transactivation and nuclear localization functions of Pichia pastoris zinc finger transcription factor Mxr1p |
title_full_unstemmed | Characterization of the transactivation and nuclear localization functions of Pichia pastoris zinc finger transcription factor Mxr1p |
title_short | Characterization of the transactivation and nuclear localization functions of Pichia pastoris zinc finger transcription factor Mxr1p |
title_sort | characterization of the transactivation and nuclear localization functions of pichia pastoris zinc finger transcription factor mxr1p |
topic | Research Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8526985/ https://www.ncbi.nlm.nih.gov/pubmed/34582889 http://dx.doi.org/10.1016/j.jbc.2021.101247 |
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