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Routray, Pratyush, Miller, J Benjamin 
ORCID: https://orcid.org/0000-0003-0882-033X, Du, Liqun, Oldroyd, Giles and Poovaiah, B W
(2013)
Phosphorylation of S344 in the calmodulin-binding domain negatively affects CCaMK function during bacterial and fungal symbioses.
The Plant Journal, 76 (2).
pp. 287-296.
ISSN 0960-7412
Abstract
Calcium and Ca(2+)/calmodulin-dependent protein kinase (CCaMK) plays a critical role in the signaling pathway that establishes root nodule symbiosis and arbuscular mycorrhizal symbiosis. Calcium-dependent autophosphorylation is central to the regulation of CCaMK, and this has been shown to promote calmodulin binding. Here, we report a regulatory mechanism of Medicago truncatula CCaMK (MtCCaMK) through autophosphorylation of S344 in the calmodulin-binding/autoinhibitory domain. The phospho-ablative mutation S344A did not have significant effect on its kinase activities, and supports root nodule symbiosis and arbuscular mycorrhizal symbiosis, indicating that phosphorylation at this position is not required for establishment of symbioses. The phospho-mimic mutation S344D show drastically reduced calmodulin-stimulated substrate phosphorylation, and this coincides with a compromised interaction with calmodulin and its interacting partner, IPD3. Functional complementation tests revealed that the S344D mutation blocked root nodule symbiosis and reduced the mycorrhizal association. Furthermore, S344D was shown to suppress the spontaneous nodulation associated with a gain-of-function mutant of MtCCaMK (T271A), revealing that phosphorylation at S344 of MtCCaMK is adequate for shutting down its activity, and is epistatic over previously identified T271 autophosphorylation. These results reveal a mechanism that enables CCaMK to 'turn off' its function through autophosphorylation.
| Item Type: | Article |
|---|---|
| Additional Information: | © 2013 The Authors The Plant Journal © 2013 John Wiley & Sons Ltd. |
| Uncontrolled Keywords: | binding sites,calcium,calcium-calmodulin-dependent protein kinases,calmodulin,genetic complementation test,medicago truncatula,mutagenesis, site-directed,mycorrhizae,phosphorylation,plant proteins,plant root nodulation,rhizobium,signal transduction,symbiosis |
| Faculty \ School: | Faculty of Science > School of Biological Sciences |
| UEA Research Groups: | Faculty of Science > Research Groups > Molecular Microbiology Faculty of Science > Research Groups > Plant Sciences |
| Depositing User: | Pure Connector |
| Date Deposited: | 04 Nov 2015 17:00 |
| Last Modified: | 25 Sep 2024 11:52 |
| URI: | https://ueaeprints.uea.ac.uk/id/eprint/54999 |
| DOI: | 10.1111/tpj.12288 |
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