2026-04-14T00:47:48Zhttps://recercat.cat/oai/request

oai:recercat.cat:2072/4888062025-11-09T13:00:36Zcom_2072_98col_2072_378192

Signaling pathways contributing to the transcriptional response to alkalinization in Komagataella phaffii : role of the transcription factors Crz1 and Rim101 Albacar, Marcel Casamayor Gracia, Antonio Zekhnini, Abdelghani Schmidt, Alexander González, Asier Ariño Carmona, Joaquín Transcriptional regulation Alkaline response Transcription factors Komagataella phaffii 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. 2025-11-09T13:00:36Z 2025-11-09T13:00:36Z 2025-11-09T13:00:36Z 2025 Article http://hdl.handle.net/2072/488806 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 Scientific reports ; Vol. 15 (2025), p. 35841 open access 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. https://creativecommons.org/licenses/by-nc-nd/4.0/