Description
Climate changes are serious challenges to agriculture and the environment. To improve plant stress tolerance the knowledge of the mechanisms of plant stress signal transduction is required. The aim of the project is elucidation of the mechanism of inhibition SnRK2 kinases (major regulators of plant stress tolerance) by their cellular partner, SCS by crystallization and solving the structure of SCS-SnRK2 complex. The results will provide information on plant stress signal transduction pathways and should be useful in the development of specific tools for the environment protection against climate changes. The obtained structure will be deposited to Protein Data Bank, presented on conferences and published in high-impact journals. The beneficent of the project will be Polish and Norwegian scientific community and in future agriculture all over the world. The bilateral relations will be strengthened by frequent communication: meetings and short-time visits of young scientists in partners’ laboratories. The Norwegian partner (group from University of Tromsø) is expert in crystallography of kinases, which is fundamental for realization of the project
Summary of project results
Salinization and desertification resulting from global climate change present serious challenges for the future of agriculture and of our environment. An understanding of the signalling pathways that control the stress tolerance of plants will give us more avenues to successfully combat negative effects of coming climate changes. The type 2 SNF1-related kinases (SnRK2s) are plant specific kinases, which play a crucial role in the regulation of plant tolerance to water deficit (drought) and salinity. Their activity is strictly controlled inside the cell by several independent mechanisms. In the laboratory of the Principal Investigator of the project, a plant specific calcium sensor that interacts with kinases studied thereby inhibiting their activity was identified. The protein was given the acronym name SCS (SnRK2-interacting calcium sensor). Our results indicate that in nature two isoforms of AtSCS exist, AtSCS-A and a shorter version of this protein, AtSCS-B. The aim of the project was elucidation of the mechanism of SnRK2’s inhibition by AtSCS-A and AtSCS-B and determination of details of the interaction between SnRK2 kinase and its inhibitor. To achieve this goal several molecular biology, biochemical, and biophysical methods have been applied, including crystallography and mass spectrometry. The results obtained during realization of the project indicate that both calcium binding proteins, AtSCS-A and AtSCS-B, interact with SnRK2s and this interaction blocks the SnRK2 activity by stabilizing the kinase activation loop in non-active conformation. We were able to identify the core of SnRK2-SCS complex. The realization of the project was possible by a cooperative effort of Polish and Norwegian partners. The results of the accomplished studies provide important insights into the signalling pathways that control plant defences in stressful environments. As demonstrated by the high priority of SnRK2 pathway studies in the agricultural sciences, the gained knowledge on the mechanism of regulation of SnRK2 activity should be important in the choice, breeding, or even design of crops and other plants to be available in response to global warming. The beneficent of the results obtained during project realization will be Polish and Norwegian scientific community and in future agriculture all over the world.
Summary of bilateral results
The realization of project was possible thanks to bilateral partnership. The objective of the project was to determine the mechanism of inhibition of SnRK2s (the kinases crucial for plant defence against harsh environmental conditions) by their specific cellular partner using various methods including analysis of protein-protein interactions, crystal structure determination and defining the core the complex responsible for the inhibition. The scientific groups, having an expertise in different research fields were responsible for different parts of the project: Polish side: Institute of Biochemistry and Biophysics, Polish Academy of Sciences, Pawińskiego 5A, Warsaw, Poland. The group of Professor Grażyna Dobrowolska was responsible for molecular biology part of the project, expression and purification of proteins studied and analysis of protein-protein interactions. Norwegian side: UiT The Arctic University of Norway, Department of Chemistry, The Norwegian Structural Biology Centre (NorStruct) , N-9037 Tromsø, Norway. Professor Richard A. Engh`s group was responsible for the crystallography part of the project, including crystallization trials, optimization, and structure solution solving. The partnership brought together experts in different research fields using different scientific techniques: Polish group - stress signal transduction in plants, including characterization, cloning, expression and purification of essential components in studied pathways, Norway group - crystallization of protein kinases with their specific effectors and solving crystal structures. Therefore, the collaboration in this consortium was highly synergistic. Partnership included sharing of materials, technologies and expertise in specific fields. The partnership also facilitated the exchange of PhD students and postdocs between Norway and Poland, that would be not possible or it would be extremely difficult without bilateral funding. We plan to extend the cooperation in framework of different international scientific programs.