The Glutamate Receptor-Like Protein GLR3.7 Interacts With 14-3-3ω and Participates in Salt Stress Response in Arabidopsis thaliana

Ionotropic glutamate receptors (iGluRs) are ligand-gated cation channels that mediate fast excitatory neurotransmission in the mammalian central nervous system. In the model plant Arabidopsis thaliana, a family of 20 glutamate receptor-like proteins (GLRs) shares similarities to animal iGluRs in seq...

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Main Authors: Po-Hsun Wang, Cheng-En Lee, Yi-Sin Lin, Man-Hsuan Lee, Pei-Yuan Chen, Hui-Chun Chang, Ing-Feng Chang
Format: Article
Language:English
Published: Frontiers Media S.A. 2019-09-01
Series:Frontiers in Plant Science
Subjects:
Online Access:https://www.frontiersin.org/article/10.3389/fpls.2019.01169/full
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spelling doaj-81ac34a5249f4debb8a6d65c875e325b2020-11-25T02:09:34ZengFrontiers Media S.A.Frontiers in Plant Science1664-462X2019-09-011010.3389/fpls.2019.01169467407The Glutamate Receptor-Like Protein GLR3.7 Interacts With 14-3-3ω and Participates in Salt Stress Response in Arabidopsis thalianaPo-Hsun Wang0Cheng-En Lee1Yi-Sin Lin2Man-Hsuan Lee3Pei-Yuan Chen4Hui-Chun Chang5Ing-Feng Chang6Ing-Feng Chang7Ing-Feng Chang8Institute of Plant Biology, National Taiwan University, Taipei, TaiwanInstitute of Plant Biology, National Taiwan University, Taipei, TaiwanInstitute of Plant Biology, National Taiwan University, Taipei, TaiwanInstitute of Plant Biology, National Taiwan University, Taipei, TaiwanInstitute of Plant Biology, National Taiwan University, Taipei, TaiwanInstitute of Plant Biology, National Taiwan University, Taipei, TaiwanInstitute of Plant Biology, National Taiwan University, Taipei, TaiwanDepartment of Life Science, National Taiwan University, Taipei, TaiwanGenome and Systems Biology Degree Program, National Taiwan University and Academia Sinica, Taipei, TaiwanIonotropic glutamate receptors (iGluRs) are ligand-gated cation channels that mediate fast excitatory neurotransmission in the mammalian central nervous system. In the model plant Arabidopsis thaliana, a family of 20 glutamate receptor-like proteins (GLRs) shares similarities to animal iGluRs in sequence and predicted secondary structure. However, the function of GLRs in plants is little known. In the present study, a serine site (Ser-860) of AtGLR3.7 phosphorylated by a calcium-dependent protein kinase (CDPK) was identified and confirmed by an in vitro kinase assay. Using a bimolecular fluorescence complementation and quartz crystal microbalance analyses, the physical interaction between AtGLR3.7 and the 14-3-3ω protein was confirmed. The mutation of Ser-860 to alanine abolished this interaction, indicating that Ser-860 is the 14-3-3ω binding site of AtGLR3.7. Compared with wild type, seed germination of the glr3.7-2 mutant was more sensitive to salt stress. However, the primary root growth of GLR3.7-S860A overexpression lines was less sensitive to salt stress than that of the wild-type line. In addition, the increase of cytosolic calcium ion concentration by salt stress was significantly lower in the glr3.7-2 mutant line than in the wild-type line. Moreover, association of 14-3-3 proteins to microsomal fractions was less in GLR3.7-S860A overexpression lines than in GLR3.7 overexpression line under 150 mM NaCl salt stress condition. Overall, our results indicated that GLR3.7 is involved in salt stress response in A. thaliana by affecting calcium signaling.https://www.frontiersin.org/article/10.3389/fpls.2019.01169/fullArabidopsisglutamate receptorkinasephosphorylation14-3-3root
collection DOAJ
language English
format Article
sources DOAJ
author Po-Hsun Wang
Cheng-En Lee
Yi-Sin Lin
Man-Hsuan Lee
Pei-Yuan Chen
Hui-Chun Chang
Ing-Feng Chang
Ing-Feng Chang
Ing-Feng Chang
spellingShingle Po-Hsun Wang
Cheng-En Lee
Yi-Sin Lin
Man-Hsuan Lee
Pei-Yuan Chen
Hui-Chun Chang
Ing-Feng Chang
Ing-Feng Chang
Ing-Feng Chang
The Glutamate Receptor-Like Protein GLR3.7 Interacts With 14-3-3ω and Participates in Salt Stress Response in Arabidopsis thaliana
Frontiers in Plant Science
Arabidopsis
glutamate receptor
kinase
phosphorylation
14-3-3
root
author_facet Po-Hsun Wang
Cheng-En Lee
Yi-Sin Lin
Man-Hsuan Lee
Pei-Yuan Chen
Hui-Chun Chang
Ing-Feng Chang
Ing-Feng Chang
Ing-Feng Chang
author_sort Po-Hsun Wang
title The Glutamate Receptor-Like Protein GLR3.7 Interacts With 14-3-3ω and Participates in Salt Stress Response in Arabidopsis thaliana
title_short The Glutamate Receptor-Like Protein GLR3.7 Interacts With 14-3-3ω and Participates in Salt Stress Response in Arabidopsis thaliana
title_full The Glutamate Receptor-Like Protein GLR3.7 Interacts With 14-3-3ω and Participates in Salt Stress Response in Arabidopsis thaliana
title_fullStr The Glutamate Receptor-Like Protein GLR3.7 Interacts With 14-3-3ω and Participates in Salt Stress Response in Arabidopsis thaliana
title_full_unstemmed The Glutamate Receptor-Like Protein GLR3.7 Interacts With 14-3-3ω and Participates in Salt Stress Response in Arabidopsis thaliana
title_sort glutamate receptor-like protein glr3.7 interacts with 14-3-3ω and participates in salt stress response in arabidopsis thaliana
publisher Frontiers Media S.A.
series Frontiers in Plant Science
issn 1664-462X
publishDate 2019-09-01
description Ionotropic glutamate receptors (iGluRs) are ligand-gated cation channels that mediate fast excitatory neurotransmission in the mammalian central nervous system. In the model plant Arabidopsis thaliana, a family of 20 glutamate receptor-like proteins (GLRs) shares similarities to animal iGluRs in sequence and predicted secondary structure. However, the function of GLRs in plants is little known. In the present study, a serine site (Ser-860) of AtGLR3.7 phosphorylated by a calcium-dependent protein kinase (CDPK) was identified and confirmed by an in vitro kinase assay. Using a bimolecular fluorescence complementation and quartz crystal microbalance analyses, the physical interaction between AtGLR3.7 and the 14-3-3ω protein was confirmed. The mutation of Ser-860 to alanine abolished this interaction, indicating that Ser-860 is the 14-3-3ω binding site of AtGLR3.7. Compared with wild type, seed germination of the glr3.7-2 mutant was more sensitive to salt stress. However, the primary root growth of GLR3.7-S860A overexpression lines was less sensitive to salt stress than that of the wild-type line. In addition, the increase of cytosolic calcium ion concentration by salt stress was significantly lower in the glr3.7-2 mutant line than in the wild-type line. Moreover, association of 14-3-3 proteins to microsomal fractions was less in GLR3.7-S860A overexpression lines than in GLR3.7 overexpression line under 150 mM NaCl salt stress condition. Overall, our results indicated that GLR3.7 is involved in salt stress response in A. thaliana by affecting calcium signaling.
topic Arabidopsis
glutamate receptor
kinase
phosphorylation
14-3-3
root
url https://www.frontiersin.org/article/10.3389/fpls.2019.01169/full
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