CDK and MAPK Synergistically Regulate Signaling Dynamics via a Shared Multi-site Phosphorylation Region on the Scaffold Protein Ste5
We report an unanticipated system of joint regulation by cyclin-dependent kinase (CDK) and mitogen-activated protein kinase (MAPK), involving collaborative multi-site phosphorylation of a single substrate. In budding yeast, the protein Ste5 controls signaling through a G1 arrest pathway. Upon cell-c...
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2018
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| Acceso en línea: | https://bibliotecadigital.exactas.uba.ar/collection/paper/document/paper_10972765_v69_n6_p938_Repetto http://hdl.handle.net/20.500.12110/paper_10972765_v69_n6_p938_Repetto |
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paper:paper_10972765_v69_n6_p938_Repetto2023-06-08T16:07:08Z CDK and MAPK Synergistically Regulate Signaling Dynamics via a Shared Multi-site Phosphorylation Region on the Scaffold Protein Ste5 Cdc28 Cks1 Cln2 cyclin G protein mating pheromone signal transduction start Ste4 cyclin dependent kinase mitogen activated protein kinase scaffold protein ste5 protein unclassified drug CLN1 protein, S cerevisiae CLN2 protein, S cerevisiae cyclin dependent kinase cycline FUS3 protein, S cerevisiae mitogen activated protein kinase protein binding Saccharomyces cerevisiae protein signal transducing adaptor protein STE5 protein, S cerevisiae Article cell cycle S phase G1 phase cell cycle checkpoint intracellular signaling MAPK signaling mass spectrometry molecular docking mutagenesis negative feedback protein binding protein electrophoresis protein phosphorylation regulatory mechanism time-lapse microscopy binding site cell cycle checkpoint cell membrane enzyme specificity enzymology genetics growth, development and aging kinetics metabolism phosphorylation Saccharomyces cerevisiae signal transduction Adaptor Proteins, Signal Transducing Binding Sites Cell Cycle Checkpoints Cell Membrane Cyclin-Dependent Kinases Cyclins Kinetics Mitogen-Activated Protein Kinases Phosphorylation Protein Binding Saccharomyces cerevisiae Saccharomyces cerevisiae Proteins Signal Transduction Substrate Specificity We report an unanticipated system of joint regulation by cyclin-dependent kinase (CDK) and mitogen-activated protein kinase (MAPK), involving collaborative multi-site phosphorylation of a single substrate. In budding yeast, the protein Ste5 controls signaling through a G1 arrest pathway. Upon cell-cycle entry, CDK inhibits Ste5 via multiple phosphorylation sites, disrupting its membrane association. Using quantitative time-lapse microscopy, we examined Ste5 membrane recruitment dynamics at different cell-cycle stages. Surprisingly, in S phase, where Ste5 recruitment should be blocked, we observed an initial recruitment followed by a steep drop-off. This delayed inhibition revealed a requirement for both CDK activity and negative feedback from the pathway MAPK Fus3. Mutagenesis, mass spectrometry, and electrophoretic analyses suggest that the CDK and MAPK modify shared sites, which are most extensively phosphorylated when both kinases are active and able to bind their docking sites on Ste5. Such collaborative phosphorylation can broaden regulatory inputs and diversify output dynamics of signaling pathways. CDKs and MAPKs phosphorylate similar sites yet generally have distinct functions and substrates. Repetto et al. uncover a case where these kinases collaborate to regulate a substrate in a signal transduction pathway by phosphorylating a shared set of sites. © 2018 Elsevier Inc. 2018 https://bibliotecadigital.exactas.uba.ar/collection/paper/document/paper_10972765_v69_n6_p938_Repetto http://hdl.handle.net/20.500.12110/paper_10972765_v69_n6_p938_Repetto |
| institution |
Universidad de Buenos Aires |
| institution_str |
I-28 |
| repository_str |
R-134 |
| collection |
Biblioteca Digital - Facultad de Ciencias Exactas y Naturales (UBA) |
| topic |
Cdc28 Cks1 Cln2 cyclin G protein mating pheromone signal transduction start Ste4 cyclin dependent kinase mitogen activated protein kinase scaffold protein ste5 protein unclassified drug CLN1 protein, S cerevisiae CLN2 protein, S cerevisiae cyclin dependent kinase cycline FUS3 protein, S cerevisiae mitogen activated protein kinase protein binding Saccharomyces cerevisiae protein signal transducing adaptor protein STE5 protein, S cerevisiae Article cell cycle S phase G1 phase cell cycle checkpoint intracellular signaling MAPK signaling mass spectrometry molecular docking mutagenesis negative feedback protein binding protein electrophoresis protein phosphorylation regulatory mechanism time-lapse microscopy binding site cell cycle checkpoint cell membrane enzyme specificity enzymology genetics growth, development and aging kinetics metabolism phosphorylation Saccharomyces cerevisiae signal transduction Adaptor Proteins, Signal Transducing Binding Sites Cell Cycle Checkpoints Cell Membrane Cyclin-Dependent Kinases Cyclins Kinetics Mitogen-Activated Protein Kinases Phosphorylation Protein Binding Saccharomyces cerevisiae Saccharomyces cerevisiae Proteins Signal Transduction Substrate Specificity |
| spellingShingle |
Cdc28 Cks1 Cln2 cyclin G protein mating pheromone signal transduction start Ste4 cyclin dependent kinase mitogen activated protein kinase scaffold protein ste5 protein unclassified drug CLN1 protein, S cerevisiae CLN2 protein, S cerevisiae cyclin dependent kinase cycline FUS3 protein, S cerevisiae mitogen activated protein kinase protein binding Saccharomyces cerevisiae protein signal transducing adaptor protein STE5 protein, S cerevisiae Article cell cycle S phase G1 phase cell cycle checkpoint intracellular signaling MAPK signaling mass spectrometry molecular docking mutagenesis negative feedback protein binding protein electrophoresis protein phosphorylation regulatory mechanism time-lapse microscopy binding site cell cycle checkpoint cell membrane enzyme specificity enzymology genetics growth, development and aging kinetics metabolism phosphorylation Saccharomyces cerevisiae signal transduction Adaptor Proteins, Signal Transducing Binding Sites Cell Cycle Checkpoints Cell Membrane Cyclin-Dependent Kinases Cyclins Kinetics Mitogen-Activated Protein Kinases Phosphorylation Protein Binding Saccharomyces cerevisiae Saccharomyces cerevisiae Proteins Signal Transduction Substrate Specificity CDK and MAPK Synergistically Regulate Signaling Dynamics via a Shared Multi-site Phosphorylation Region on the Scaffold Protein Ste5 |
| topic_facet |
Cdc28 Cks1 Cln2 cyclin G protein mating pheromone signal transduction start Ste4 cyclin dependent kinase mitogen activated protein kinase scaffold protein ste5 protein unclassified drug CLN1 protein, S cerevisiae CLN2 protein, S cerevisiae cyclin dependent kinase cycline FUS3 protein, S cerevisiae mitogen activated protein kinase protein binding Saccharomyces cerevisiae protein signal transducing adaptor protein STE5 protein, S cerevisiae Article cell cycle S phase G1 phase cell cycle checkpoint intracellular signaling MAPK signaling mass spectrometry molecular docking mutagenesis negative feedback protein binding protein electrophoresis protein phosphorylation regulatory mechanism time-lapse microscopy binding site cell cycle checkpoint cell membrane enzyme specificity enzymology genetics growth, development and aging kinetics metabolism phosphorylation Saccharomyces cerevisiae signal transduction Adaptor Proteins, Signal Transducing Binding Sites Cell Cycle Checkpoints Cell Membrane Cyclin-Dependent Kinases Cyclins Kinetics Mitogen-Activated Protein Kinases Phosphorylation Protein Binding Saccharomyces cerevisiae Saccharomyces cerevisiae Proteins Signal Transduction Substrate Specificity |
| description |
We report an unanticipated system of joint regulation by cyclin-dependent kinase (CDK) and mitogen-activated protein kinase (MAPK), involving collaborative multi-site phosphorylation of a single substrate. In budding yeast, the protein Ste5 controls signaling through a G1 arrest pathway. Upon cell-cycle entry, CDK inhibits Ste5 via multiple phosphorylation sites, disrupting its membrane association. Using quantitative time-lapse microscopy, we examined Ste5 membrane recruitment dynamics at different cell-cycle stages. Surprisingly, in S phase, where Ste5 recruitment should be blocked, we observed an initial recruitment followed by a steep drop-off. This delayed inhibition revealed a requirement for both CDK activity and negative feedback from the pathway MAPK Fus3. Mutagenesis, mass spectrometry, and electrophoretic analyses suggest that the CDK and MAPK modify shared sites, which are most extensively phosphorylated when both kinases are active and able to bind their docking sites on Ste5. Such collaborative phosphorylation can broaden regulatory inputs and diversify output dynamics of signaling pathways. CDKs and MAPKs phosphorylate similar sites yet generally have distinct functions and substrates. Repetto et al. uncover a case where these kinases collaborate to regulate a substrate in a signal transduction pathway by phosphorylating a shared set of sites. © 2018 Elsevier Inc. |
| title |
CDK and MAPK Synergistically Regulate Signaling Dynamics via a Shared Multi-site Phosphorylation Region on the Scaffold Protein Ste5 |
| title_short |
CDK and MAPK Synergistically Regulate Signaling Dynamics via a Shared Multi-site Phosphorylation Region on the Scaffold Protein Ste5 |
| title_full |
CDK and MAPK Synergistically Regulate Signaling Dynamics via a Shared Multi-site Phosphorylation Region on the Scaffold Protein Ste5 |
| title_fullStr |
CDK and MAPK Synergistically Regulate Signaling Dynamics via a Shared Multi-site Phosphorylation Region on the Scaffold Protein Ste5 |
| title_full_unstemmed |
CDK and MAPK Synergistically Regulate Signaling Dynamics via a Shared Multi-site Phosphorylation Region on the Scaffold Protein Ste5 |
| title_sort |
cdk and mapk synergistically regulate signaling dynamics via a shared multi-site phosphorylation region on the scaffold protein ste5 |
| publishDate |
2018 |
| url |
https://bibliotecadigital.exactas.uba.ar/collection/paper/document/paper_10972765_v69_n6_p938_Repetto http://hdl.handle.net/20.500.12110/paper_10972765_v69_n6_p938_Repetto |
| _version_ |
1768543243328290816 |