Direct interaction of a divergent CaM isoform and the transcription factor, MYB2, enhances salt tolerance in arabidopsis.
Calmodulin (CaM), a ubiquitous calcium-binding protein, regulates diverse cellular functions by modulating the activity of a variety of enzymes and proteins. Plants express numerous CaM isoforms that exhibit differential activation and/or inhibition of CaM-dependent enzymes in vitro. However, the specific biological functions of plant CaM are not well known. In this study, we isolated a cDNA encoding a CaM binding transcription factor, MYB2, that regulates the expression of salt- and dehydration-responsive genes in Arabidopsis. This was achieved using a salt-inducible CaM isoform (GmCaM4) as a probe from a salt-treated Arabidopsis expression library. Using domain mapping, we identified a Ca2+-dependent CaM binding domain in MYB2. The specific binding of CaM to CaM binding domain was confirmed by site-directed mutagenesis, a gel mobility shift assay, split ubiquitin assay, and a competition assay using a Ca2+/CaM-dependent enzyme. Interestingly, the specific CaM isoform GmCaM4 enhances the DNA binding activity of AtMYB2, whereas this was inhibited by a closely related CaM isoform (GmCaM1). Overexpression of Gm-CaM4 in Arabidopsis up-regulates the transcription rate of AtMYB2-regulated genes, including the proline-synthesizing enzyme P5CS1 (Delta1-pyrroline-5-carboxylate synthetase-1), which confers salt tolerance by facilitating proline accumulation. Therefore, we suggest that a specific CaM isoform mediates salt-induced Ca2+ signaling through the activation of an MYB transcriptional activator, thereby resulting in salt tolerance in plants.
Yoo
Jae Hyuk
JH
Division of Applied Life Science (BK21 Program), Plant Molecular Biology and Biotechnology Research Center, and Environmental Biotechnology National Core Research Center, Gyeongsang National University, Jinju 660-701, Korea.
Park
Chan Young
CY
Kim
Jong Cheol
JC
Heo
Won Do
WD
Cheong
Mi Sun
MS
Park
Hyeong Cheol
HC
Kim
Min Chul
MC
Moon
Byeong Cheol
BC
Choi
Man Soo
MS
Kang
Yun Hwan
YH
Lee
Ju Huck
JH
Kim
Ho Soo
HS
Lee
Sang Min
SM
Yoon
Hae Won
HW
Lim
Chae Oh
CO
Yun
Dae-Jin
DJ
Lee
Sang Yeol
SY
Chung
Woo Sik
WS
Cho
Moo Je
MJ
D017360:Arabidopsis / Q000235:genetics / Q000378:metabolism*; D029681:Arabidopsis Proteins / Q000235:genetics / Q000378:metabolism*; D001665:Binding Sites / Q000235:genetics; D020013:Calcium Signaling / Q000502:physiology; D002147:Calmodulin / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism*; D018076:DNA, Complementary; D018506:Gene Expression Regulation, Plant / Q000502:physiology; D007536:Isomerism; D016297:Mutagenesis, Site-Directed; D030821:Plants, Genetically Modified; D011392:Proline / Q000378:metabolism; D012492:Salts / Q000378:metabolism*; D015534:Trans-Activators / Q000235:genetics / Q000378:metabolism*; D025801:Ubiquitin; D015003:Yeasts
D029681:Arabidopsis Proteins; D002147:Calmodulin; D018076:DNA, Complementary; C104175:MYB2 protein, Arabidopsis; D012492:Salts; D015534:Trans-Activators; D025801:Ubiquitin; D011392:Proline