Direct Interaction of a Divergent CaM Isoform and the Transcription Factor, MYB2, Enhances Salt Tolerance in Arabidopsis

• Published in: The Journal of biological chemistry Vol. 280; no. 5; pp. 3697 - 3706
• Main Authors: Yoo, Jae Hyuk, Park, Chan Young, Kim, Jong Cheol, Do Heo, Won, Cheong, Mi Sun, Park, Hyeong Cheol, Kim, Min Chul, Moon, Byeong Cheol, Choi, Man Soo, Kang, Yun Hwan, Lee, Ju Huck, Kim, Ho Soo, Lee, Sang Min, Yoon, Hae Won, Lim, Chae Oh, Yun, Dae-Jin, Lee, Sang Yeol, Chung, Woo Sik, Cho, Moo Je
• Format: Journal Article
• Language: English
• Published: United States Elsevier Inc 04-02-2005; American Society for Biochemistry and Molecular Biology
• Subjects: Arabidopsis; Arabidopsis - genetics; Arabidopsis - metabolism; Arabidopsis Proteins - genetics; Arabidopsis Proteins - metabolism; Binding Sites - genetics; Calcium Signaling - physiology; Calmodulin - chemistry; Calmodulin - genetics; Calmodulin - metabolism; DNA, Complementary; Gene Expression Regulation, Plant - physiology; Isomerism; Mutagenesis, Site-Directed; Plants, Genetically Modified; Proline - metabolism; Salts - metabolism; Trans-Activators - genetics; Trans-Activators - metabolism; Ubiquitin; Yeasts
• ISSN: 0021-9258; 1083-351X
• DOI: 10.1074/jbc.M408237200

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 (Δ1-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.