Cargando…

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...

Descripción completa

Detalles Bibliográficos
Autores principales: Gupta, Aditi, Krishna Rao, Kamisetty, Sahu, Umakant, Rangarajan, Pundi N.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: American Society for Biochemistry and Molecular Biology 2021
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
_version_ 1784585983268749312
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
work_keys_str_mv AT guptaaditi characterizationofthetransactivationandnuclearlocalizationfunctionsofpichiapastoriszincfingertranscriptionfactormxr1p
AT krishnaraokamisetty characterizationofthetransactivationandnuclearlocalizationfunctionsofpichiapastoriszincfingertranscriptionfactormxr1p
AT sahuumakant characterizationofthetransactivationandnuclearlocalizationfunctionsofpichiapastoriszincfingertranscriptionfactormxr1p
AT rangarajanpundin characterizationofthetransactivationandnuclearlocalizationfunctionsofpichiapastoriszincfingertranscriptionfactormxr1p