FRET kinase sensor development reveals SnRK2/OST1 activation by ABA but not by MeJA and high CO2 during stomatal closure
Sucrose-non-fermenting-1-related protein kinase-2s (SnRK2s) are critical for plant abiotic stress responses, including abscisic acid (ABA) signaling. Here, we develop a genetically encoded reporter for SnRK2 kinase activity. This sensor, named SNACS, shows an increase in the ratio of yellow to cyan...
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| Format: | Article |
| Language: | English |
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eLife Sciences Publications Ltd
2020-05-01
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| Series: | eLife |
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| Online Access: | https://elifesciences.org/articles/56351 |
| _version_ | 1818019641730007040 |
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| author | Li Zhang Yohei Takahashi Po-Kai Hsu Hannes Kollist Ebe Merilo Patrick J Krysan Julian I Schroeder |
| author_facet | Li Zhang Yohei Takahashi Po-Kai Hsu Hannes Kollist Ebe Merilo Patrick J Krysan Julian I Schroeder |
| author_sort | Li Zhang |
| collection | DOAJ |
| description | Sucrose-non-fermenting-1-related protein kinase-2s (SnRK2s) are critical for plant abiotic stress responses, including abscisic acid (ABA) signaling. Here, we develop a genetically encoded reporter for SnRK2 kinase activity. This sensor, named SNACS, shows an increase in the ratio of yellow to cyan fluorescence emission by OST1/SnRK2.6-mediated phosphorylation of a defined serine residue in SNACS. ABA rapidly increases FRET efficiency in N. benthamiana leaf cells and Arabidopsis guard cells. Interestingly, protein kinase inhibition decreases FRET efficiency in guard cells, providing direct experimental evidence that basal SnRK2 activity prevails in guard cells. Moreover, in contrast to ABA, the stomatal closing stimuli, elevated CO2 and MeJA, did not increase SNACS FRET ratios. These findings and gas exchange analyses of quintuple/sextuple ABA receptor mutants show that stomatal CO2 signaling requires basal ABA and SnRK2 signaling, but not SnRK2 activation. A recent model that CO2 signaling is mediated by PYL4/PYL5 ABA-receptors could not be supported here in two independent labs. We report a potent approach for real-time live-cell investigations of stress signaling. |
| first_indexed | 2024-04-14T07:53:54Z |
| format | Article |
| id | doaj.art-889e1473a5bc4126ab12154567e993e9 |
| institution | Directory Open Access Journal |
| issn | 2050-084X |
| language | English |
| last_indexed | 2024-04-14T07:53:54Z |
| publishDate | 2020-05-01 |
| publisher | eLife Sciences Publications Ltd |
| record_format | Article |
| series | eLife |
| spelling | doaj.art-889e1473a5bc4126ab12154567e993e92022-12-22T02:05:06ZengeLife Sciences Publications LtdeLife2050-084X2020-05-01910.7554/eLife.56351FRET kinase sensor development reveals SnRK2/OST1 activation by ABA but not by MeJA and high CO2 during stomatal closureLi Zhang0https://orcid.org/0000-0003-0467-0290Yohei Takahashi1https://orcid.org/0000-0002-9406-4093Po-Kai Hsu2https://orcid.org/0000-0001-7265-7077Hannes Kollist3Ebe Merilo4Patrick J Krysan5Julian I Schroeder6https://orcid.org/0000-0002-3283-5972Cell and Developmental Biology Section, Division of Biological Sciences, University of California, San Diego, San Diego, United StatesCell and Developmental Biology Section, Division of Biological Sciences, University of California, San Diego, San Diego, United StatesCell and Developmental Biology Section, Division of Biological Sciences, University of California, San Diego, San Diego, United StatesInstitute of Technology, University of Tartu, Tartu, EstoniaInstitute of Technology, University of Tartu, Tartu, EstoniaHorticulture Department, University of Wisconsin-Madison, Madison, United StatesCell and Developmental Biology Section, Division of Biological Sciences, University of California, San Diego, San Diego, United StatesSucrose-non-fermenting-1-related protein kinase-2s (SnRK2s) are critical for plant abiotic stress responses, including abscisic acid (ABA) signaling. Here, we develop a genetically encoded reporter for SnRK2 kinase activity. This sensor, named SNACS, shows an increase in the ratio of yellow to cyan fluorescence emission by OST1/SnRK2.6-mediated phosphorylation of a defined serine residue in SNACS. ABA rapidly increases FRET efficiency in N. benthamiana leaf cells and Arabidopsis guard cells. Interestingly, protein kinase inhibition decreases FRET efficiency in guard cells, providing direct experimental evidence that basal SnRK2 activity prevails in guard cells. Moreover, in contrast to ABA, the stomatal closing stimuli, elevated CO2 and MeJA, did not increase SNACS FRET ratios. These findings and gas exchange analyses of quintuple/sextuple ABA receptor mutants show that stomatal CO2 signaling requires basal ABA and SnRK2 signaling, but not SnRK2 activation. A recent model that CO2 signaling is mediated by PYL4/PYL5 ABA-receptors could not be supported here in two independent labs. We report a potent approach for real-time live-cell investigations of stress signaling.https://elifesciences.org/articles/56351SnRK2 OST1 protein kinasestomatal guard cellsin vivo FRET imagingabscisic acid14-3-3 proteinplant stress signaling |
| spellingShingle | Li Zhang Yohei Takahashi Po-Kai Hsu Hannes Kollist Ebe Merilo Patrick J Krysan Julian I Schroeder FRET kinase sensor development reveals SnRK2/OST1 activation by ABA but not by MeJA and high CO2 during stomatal closure eLife SnRK2 OST1 protein kinase stomatal guard cells in vivo FRET imaging abscisic acid 14-3-3 protein plant stress signaling |
| title | FRET kinase sensor development reveals SnRK2/OST1 activation by ABA but not by MeJA and high CO2 during stomatal closure |
| title_full | FRET kinase sensor development reveals SnRK2/OST1 activation by ABA but not by MeJA and high CO2 during stomatal closure |
| title_fullStr | FRET kinase sensor development reveals SnRK2/OST1 activation by ABA but not by MeJA and high CO2 during stomatal closure |
| title_full_unstemmed | FRET kinase sensor development reveals SnRK2/OST1 activation by ABA but not by MeJA and high CO2 during stomatal closure |
| title_short | FRET kinase sensor development reveals SnRK2/OST1 activation by ABA but not by MeJA and high CO2 during stomatal closure |
| title_sort | fret kinase sensor development reveals snrk2 ost1 activation by aba but not by meja and high co2 during stomatal closure |
| topic | SnRK2 OST1 protein kinase stomatal guard cells in vivo FRET imaging abscisic acid 14-3-3 protein plant stress signaling |
| url | https://elifesciences.org/articles/56351 |
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