Drafting Penicillium oxalicum calcineurin-CrzA pathway by combining the analysis of phenotype, transcriptome, and
Kaili Zhao1, Zhongjiao Liu1, Mengxue Li1
1National Glycoengineering Research Center, Shandong University, No. 72 Binhai Road, Qingdao 266237, China.
Abstract:
Fungi sense environmental signals and coordinate growth, development, and metabolism accordingly. Calcium-calmodulin-calcineurin signaling is a conserved cascade pathway in fungi. One of the most important downstream targets of this pathway is the transcription factor Crz1/CrzA, which plays an essential role in various cellular processes. The putative collaborators of Penicillium oxalicum CrzA (PoCrzA) were found, through tandem affinity purification followed by mass spectrometric analysis (TAP-MS). A total of 50 protein-protein interaction collaborators of PoCrzA were observed. Among them, some collaborators, such as the catalytic subunit of calcineurin (Cna1, calcineurin A), the regulatory catalytic subunit of calcineurin (Cnb1, calcineurin B), and a 14-3-3 protein Bmh1, which were previously reported in yeast, were identified. Some putative collaborators, including two karyopherins (exportin Los1 and importin Srp1), two kinases (Fus3 and Slt2p), and a general transcriptional corepressor (Cyc8), were also found. The CrzA deletion mutant ΔPocrzA exhibited slow hyphal growth, impaired conidiogenesis, and reduced extracellular cellulase synthesis. Phenotype and transcriptome analysis showed that PoCrzA regulated fungal development in a Flbs-BrlA-dependent manner and participated in cellulase synthesis by modulating cellulolytic gene expression. On the basis of the results of TAP-MS, transcriptome, and phenotypic analysis in P. oxalicum, our study was the first to draft the calcineurin-CrzA pathway in cellulolytic fungi.
Insights
The calcineurin-CrzA pathway in Penicillium oxalicum is crucial for fungal development and cellulase production. This study identifies key protein interactions and regulatory mechanisms, advancing our understanding of fungal signaling.
Area of Science:
- Mycology
- Molecular Biology
- Biochemistry
Background:
- Fungi utilize conserved signaling pathways, like calcium-calmodulin-calcineurin, to respond to environmental cues.
- The transcription factor Crz1/CrzA is a key downstream effector in this pathway, regulating vital cellular processes.
Purpose of the Study:
- To identify protein-protein interactions of Penicillium oxalicum CrzA (PoCrzA) using TAP-MS.
- To elucidate the role of the calcineurin-CrzA pathway in fungal development and cellulase synthesis in P. oxalicum.
Main Methods:
- Tandem affinity purification followed by mass spectrometry (TAP-MS) to identify PoCrzA interactors.
- Phenotypic analysis of a ΔPocrzA deletion mutant.
- Transcriptome analysis to understand gene expression regulation.
Main Results:
- Identified 50 protein-protein interaction partners of PoCrzA, including known components like calcineurin subunits (Cna1, Cnb1) and 14-3-3 protein (Bmh1), as well as novel interactors (karyopherins, kinases, corepressor).
- The ΔPocrzA mutant showed defects in hyphal growth, conidiogenesis, and cellulase synthesis.
- PoCrzA regulates fungal development via the Flbs-BrlA pathway and modulates cellulolytic gene expression.
Conclusions:
- The study provides the first draft of the calcineurin-CrzA pathway in cellulolytic fungi.
- PoCrzA is a critical regulator of both fungal development and the production of extracellular cellulases in P. oxalicum.
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