The Small Yeast GTPase Rho5 and Its Dimeric GEF Dck1/Lmo1 Respond to Glucose Starvation
Abstract
:1. Introduction
2. Results
2.1. Dck1 Forms Transient Foci in the Cytoplasm and Rapidly Accumulates at Mitochondria upon Glucose Starvation
2.2. Rho5 Deletions Genetically Interact with Gpr1, Gpa2, and Sch9
2.3. Deletions of RHO5, GPR1, GPA2, or RAS2 Result in Hyper-Resistance towards Cell Wall Stress Agents
2.4. Lack of Dck1 Moderately Affects Actin Dynamics in Budding Yeast Cells
3. Discussion
4. Materials and Methods
4.1. Strains and Growth Conditions
4.2. Construction of Plasmids, Deletion Mutants and Gene Tagging
4.3. Strains and Experimental Conditions for Fluorescence Microscopy, Image Acquisition and Statistical Analyses
Supplementary Materials
Author Contributions
Funding
Acknowledgments
Conflicts of Interest
Abbreviations
AMPK | 5′ adenosine monophosphate activated kinase |
2-DOG | 2-deoxy glucose |
GEF | GDP/GTP exchange factor |
PKA | protein kinase A |
ROS | reactive oxygen species |
SNF | sucrose non-fermenters |
TORC1 | target of rapamycin complex 1 |
vATPase | vacuolar ATPase complex |
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Strain | Genotype | Source |
---|---|---|
DHD5 | MATa/α ura3-52/ura3-52 leu2-3,112/leu2-3,112 his3-11,15/his3-11,15 | [47] |
HAJ6-A | MATa segregant from DHD5 | [50] |
HAJ6-B | MATα segregant from DHD5 | [50] |
DAJ119 | as DHD5 except dck1::kanMX/DCK1 sch9::SkHIS3/SCH9 | This study |
DAJ128 | as DHD5 except lmo1::kanMX/LMO1 sch9::SkHIS3/SCH9 | This study |
DAJ138 | as DHD5 except rho5::KanMX/RHO5 sch9::SkHIS3/SCH9 | This study |
DAJ139 | as DHD5 except ras1::KanMX /RAS1 sch9::SkHIS3/SCH9 | This study |
DAJ140 | as DHD5 except ras2::KanMX /RAS2 sch9::SkHIS3/SCH9 | This study |
DAJ144 | as DHD5 except rho5::KanMX/RHO5 gpa2::KanMX/GPA2 | This study |
DAJ145 | as DHD5 except rho5::KanMX/RHO5 gpr1::KanMX/GPR1 | This study |
HAJ152-A | as HAJ6-A except DCK1-3xmyEGFP::SpHIS5 | [10] |
HAJ187-A | as HAJ6-A except gpr1::KanMX | This study |
HAJ188-A | as HAJ6-A except gpa2::KanMX | This study |
HAJ201-A | as HAJ6-A except lmo1::KanMX | This study |
HAJ201-B | as HAJ6-B except lmo1::KanMX | This study |
HAJ204-A | as HAJ6-A except gpr1::KanMX DCK1-3xEGFP::SpHIS5 | This study |
HAJ205-A | as HAJ6-A except gpa2::KanMX DCK1-3xEGFP::SpHIS5 | This study |
HAJ206-A | as HAJ6-A except sch9::KanMX DCK1-3xEGFP::SpHIS5 | This study |
HAJ207-A | as HAJ6-A except lmo1::KanMX DCK1-3xEGFP::SpHIS5 | This study |
HAJ216-A | as HAJ6-B except rho5::KanMX | [10] |
HAJ217-A | as HAJ6-A except ras1::KanMX | This study |
HAJ218-A | as HAJ6-A except ras2::KanMX | This study |
HCS076-A | as HAJ6-A except rho5::KanMX IDP1-mCherry::SkHIS3 | This study |
HMZ18-A | as HAJ6-A except lmo1::SkHIS3 | This study |
HOD294.2 | as DHD5 except rho5::kanMX/RHO5 snf1::SpHIS5/SNF1 | This study |
HOD320 | as DHD5 except rho5::kanMX/RHO5 ras2::SkHIS3/RAS2 | This study |
HOD294.2-1A | as HAJ6-A | This study |
HOD294.2-1B | as HAJ6-B | This study |
HOD294.2-2B | as HAJ6-B except rho5::KanMX | This study |
HOD294.2-3B | as HAJ6-A except rho5::KanMX | This study |
HOD309-1D | as HAJ6-A except rho5::KanMX CAP2-EGFP::SpHIS5 | This study |
HOD310-1B | as HAJ6-A except CAP2-EGFP::SpHIS5 | This study |
HOD310-4A | as HAJ6-A except lmo1::SkHIS3 CAP2-EGFP::SpHIS5 | This study |
HOD314-2C | as HAJ6-A except dck1::KlURA3 CAP2-EGFP::SpHIS5 | This study |
HOD314-5A | as HAJ6-A except dck1::KlURA3 | This study |
HOD314-8A | as HAJ6-B except dck1::KlURA3 | This study |
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Schmitz, H.-P.; Jendretzki, A.; Sterk, C.; Heinisch, J.J. The Small Yeast GTPase Rho5 and Its Dimeric GEF Dck1/Lmo1 Respond to Glucose Starvation. Int. J. Mol. Sci. 2018, 19, 2186. https://doi.org/10.3390/ijms19082186
Schmitz H-P, Jendretzki A, Sterk C, Heinisch JJ. The Small Yeast GTPase Rho5 and Its Dimeric GEF Dck1/Lmo1 Respond to Glucose Starvation. International Journal of Molecular Sciences. 2018; 19(8):2186. https://doi.org/10.3390/ijms19082186
Chicago/Turabian StyleSchmitz, Hans-Peter, Arne Jendretzki, Carolin Sterk, and Jürgen J. Heinisch. 2018. "The Small Yeast GTPase Rho5 and Its Dimeric GEF Dck1/Lmo1 Respond to Glucose Starvation" International Journal of Molecular Sciences 19, no. 8: 2186. https://doi.org/10.3390/ijms19082186