Protein Phosphatase Ppz1 Is Not Regulated by a Hal3-Like Protein in Plant Pathogen Ustilago maydis
Abstract
:1. Introduction
2. Results
2.1. Identification of Ustilago maydis Ppz1 and Hal3-Like Proteins
2.2. Expression and Functional Characterization of UmPpz1 in S. cerevisiae
2.3. Functional Characterization of UmHal3 in S. cerevisiae
2.4. Characterization of Recombinant UmPpz1 and UmHal3 Proteins
2.5. UmHal3 Is a Functional PPCDC Enzyme
2.6. Phenotypic Characterization of UmPpz1 and UmHal3 Deletion Mutants
2.7. Ppz1 Is Dispensable for Virulence in U. maydis
3. Discussion
4. Materials and Methods
4.1. Yeast Strains and Growth Conditions
4.2. Generation of Fungal Deletion Mutant Strains
4.3. Recombinant DNA Techniques and Plasmid Construction
4.4. Phenotypic Analysis
4.5. Expression and Purification of Recombinant Proteins
4.6. Yeasts Protein Extraction and Immunoblot Analysis
4.7. In Vitro Interaction Experiments
4.8. Other Techniques
Supplementary Materials
Author Contributions
Funding
Acknowledgments
Conflicts of Interest
References
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Organism | Strain | Genotype | Source/Reference |
---|---|---|---|
U. maydis | |||
FB1 | a1 b1 | [35] | |
FB2 | a2 b2 | [35] | |
FBD11 | a1a2 b1b2 | [35] | |
UMP341 | a1a2 b1b2 hal3Δ::HygR/hal3 | This work | |
UMP342 | a1a2 b1b2 ppz1Δ::HygR/ppz1 | This work | |
UMP351 | a1 b1 ppz1Δ::HygR | This work | |
UMP352 | a2 b2 ppz1Δ::HygR | This work | |
S. cerevisiae | |||
BY4741 | MATa his3Δ1 leu2Δ0 met15Δ0 ura3Δ0 | Euroscarf | |
ppz1 | BY4741 ppz1::kanMx4 | Euroscarf | |
hal3 | BY4741 hal3::kanMx4 | Euroscarf | |
slt2 | BY4741 slt2::kanMx4 | Euroscarf | |
IM21 | JA100 (MATa, ura3-52 leu2-3,112 trp1-1 his4 can-1r) ppz1::kanMx4 hal3::LEU2 | [42] | |
AGS04 | 1788 (a/α, ura3-52 leu2-3,112 trp1-1 his4 can-1r) VHS3/vhs3::kanMx4 HAL3/hal3::LEU2 | [15] | |
MAR25 | 1788 (a/α, ura3-52 leu2-3,112 trp1-1 his4 can-1r) CAB3/cab3::kanMx4 | [25] |
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Zhang, C.; de la Torre, A.; Pérez-Martín, J.; Ariño, J. Protein Phosphatase Ppz1 Is Not Regulated by a Hal3-Like Protein in Plant Pathogen Ustilago maydis. Int. J. Mol. Sci. 2019, 20, 3817. https://doi.org/10.3390/ijms20153817
Zhang C, de la Torre A, Pérez-Martín J, Ariño J. Protein Phosphatase Ppz1 Is Not Regulated by a Hal3-Like Protein in Plant Pathogen Ustilago maydis. International Journal of Molecular Sciences. 2019; 20(15):3817. https://doi.org/10.3390/ijms20153817
Chicago/Turabian StyleZhang, Chunyi, Antonio de la Torre, José Pérez-Martín, and Joaquín Ariño. 2019. "Protein Phosphatase Ppz1 Is Not Regulated by a Hal3-Like Protein in Plant Pathogen Ustilago maydis" International Journal of Molecular Sciences 20, no. 15: 3817. https://doi.org/10.3390/ijms20153817