Signaling pathways contributing to the transcriptional response to alkalinization in Komagataella phaffii : role of the transcription factors Crz1 and Rim101
Albacar, Marcel (Universitat Autònoma de Barcelona. Institut de Biotecnologia i de Biomedicina "Vicent Villar Palasí")
Casamayor Gracia, Antonio (Universitat Autònoma de Barcelona. Departament de Bioquímica i de Biologia Molecular)
Zekhnini, Abdelghani
Schmidt, Alexander
Gonzalez Sevine, Asier (Universitat Autònoma de Barcelona. Departament de Bioquímica i de Biologia Molecular)
Ariño Carmona, Joaquín (Universitat Autònoma de Barcelona. Departament de Bioquímica i de Biologia Molecular)

Date:	2025
Description:	18 pàg.
Abstract:	Promoters able to respond to moderate alkalinization in the industrially relevant yeast Komagataella phaffii have been proposed as a tool for the design of novel methanol-free platforms for heterologous protein expression. However, the molecular bases of such response are unknown. We identify here the transcription factors Crz1 and Rim101 in K. phaffii and use CRISPR/Cas9 techniques to create single and double mutant strains. While crz1 cells are strongly sensitive to Ca2+ ions, mildly sensitive to alkaline pH and hypertolerant to SDS, rim101 cells are markedly hypersensitive to Na+ and Li+ ions, to alkaline pH and to SDS. RNA-seq analysis revealed that mutation of RIM101 affects 55% of alkaline pH-responsive genes, whereas lack of Crz1 alters only 11% of these. Thirty-eight genes are co-regulated by both Crz1 and Rim101. The PMC1 vacuolar Ca2+-ATPase is regulated by Crz1, whereas the ENA2 Na+-ATPase, the PHO89 and PHO84 phosphate transporters, and diverse genes involved in iron homeostasis are Rim101-dependent. These effects were confirmed by proteomic analyses after 3 h of alkalinization. Therefore, Rim101 appears as a major player in the transcriptional response to alkalinization and, together with Crz1, they account for nearly 60% of the alkaline pH transcriptional response in K. phaffii.
Grants:	Agència de Gestió d'Ajuts Universitaris i de Recerca 2023/PROD-00006 Agencia Estatal de Investigación PID2020-113319RB-I00 Agencia Estatal de Investigación PID2023-150535OB-I00
Rights:	Aquest document està subjecte a una llicència d'ús Creative Commons. Es permet la reproducció total o parcial, la distribució, i la comunicació pública de l'obra, sempre que no sigui amb finalitats comercials, i sempre que es reconegui l'autoria de l'obra original. No es permet la creació d'obres derivades.
Language:	Anglès
Document:	Article ; recerca ; Versió publicada
Subject:	Transcriptional regulation ; Alkaline response ; Transcription factors ; Komagataella phaffii
Published in:	Scientific reports, Vol. 15 (2025) , p. 35841, ISSN 2045-2322

DOI: 10.1038/s41598-025-20192-6
PMID: 41087568

18 p, 4.4 MB

The record appears in these collections:
Research literature > UAB research groups literature > Research Centres and Groups (research output) > Health sciences and biosciences > Institut de Biotecnologia i de Biomedicina (IBB)
Articles > Research articles
Articles > Published articles