Object's details: SNF1-RELATED PROTEIN KINASES 2 ARE NEGATIVELY REGULATED BY A PLANT-SPECIFIC CALCIUM SENSOR

Provider:Repozytorium Instytutu Biochemii i Biofizyki Polskiej Akademii Nauk

Description

Title:
SNF1-RELATED PROTEIN KINASES 2 ARE NEGATIVELY REGULATED BY A PLANT-SPECIFIC CALCIUM SENSOR
Creator:
Bucholc, Maria;
Ciesielski, Arkadiusz;
Goch, Grażyna;
Anielska-Mazur, Anna;
Kulik, Anna;
Krzywińska, Ewa;
Dobrowolska, Grażyna
Description:
SNF1-related protein kinases 2 (SnRK2s) are plant-specific enzymes involved in environmental stress signaling and abscisic acid-regulated plant development. Here, we report that SnRK2s interact with and are regulated by a plant-specific calcium-binding protein. We screened a Nicotiana plumbaginifolia Matchmaker cDNA library for proteins interacting with Nicotiana tabacum osmotic stress-activated protein kinase (NtOSAK), a member of the SnRK2 family. A putative EF-hand calcium-binding protein was identified as a molecular partner of NtOSAK. To determine whether the identified protein interacts only with NtOSAK or with other SnRK2s as well, we studied interaction of an Arabidopsis thaliana orthologue of the calcium-binding protein with selected Arabidopsis SnRK2s using a two-hybrid system. All kinases studied interacted with the protein. The interactions were confirmed by bimolecular fluorescence complementation assay, indicating that the binding occurs in planta, exclusively in the cytoplasm. Calcium binding properties of the protein were analyzed by fluorescence spectroscopy using Tb3+ as a spectroscopic probe. The calcium binding constant, determined by the protein fluorescence titration was 2.5±0.9x105 M-1. The CD spectrum indicated that the secondary structure of the protein changes significantly in the presence of calcium, suggesting its possible function as a calcium sensor in plant cells. In vitro studies revealed that the activity of SnRK2 kinases analyzed is inhibited in a calcium-dependent manner by the identified calcium sensor which we named SCS (SnRK2-interacting Calcium Sensor). Our results suggest that SCS is involved in response to ABA during seed germination most probably by a negative regulation of SnRK2s activity.
Object availability:
free access
Date:
2011
Type:
Article;
PeerReviewed
Language:
English
Format:
application/pdf
Relation: http://eprints.ibb.waw.pl/74
Subject:
QK Botany
Identifier: http://eprints.ibb.waw.pl/74/1/Bucholc_et_al_manuscript_18_Nov_2010.doc.pdf ;
Bucholc, Maria and Ciesielski, Arkadiusz and Goch, Grażyna and Anielska-Mazur, Anna and Kulik, Anna and Krzywińska, Ewa and Dobrowolska, Grażyna (2011) SNF1-RELATED PROTEIN KINASES 2 ARE NEGATIVELY REGULATED BY A PLANT-SPECIFIC CALCIUM SENSOR. Journal of Biological Chemistry, 286 (5). pp. 3429-3441.
Data provider:
Repozytorium Instytutu Biochemii i Biofizyki Polskiej Akademii Nauk

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Object's details: SNF1-RELATED PROTEIN KINASES 2 ARE NEGATIVELY REGULATED BY A PLANT-SPECIFIC CALCIUM SENSOR

Description

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