AtKC1 and CIPK23 Synergistically Modulate AKT1-Mediated Low-Potassium Stress Responses in Arabidopsis

• Published in: Plant physiology (Bethesda) 2016-04, Vol.170 (4), p.2264-2277
• Main Authors: Wang, Xue-Ping, Chen, Li-Mei, Liu, Wen-Xin, Shen, Li-Ke, Wang, Feng-Liu, Zhou, Yuan, Zhang, Ziding, Wu, Wei-Hua, Wang, Yi
• Format: Article
• Language: English
• Subjects: Amino Acid Sequence; Arabidopsis - drug effects; Arabidopsis - physiology; Arabidopsis Proteins - chemistry; Arabidopsis Proteins - genetics; Arabidopsis Proteins - metabolism; Cloning, Molecular; Ethyl Methanesulfonate; Genetic Complementation Test; Germination - drug effects; MEMBRANES, TRANSPORT AND BIOENERGETICS; Mutagenesis; Mutation - genetics; Phenotype; Plant Shoots - drug effects; Plant Shoots - metabolism; Potassium - pharmacology; Potassium Channels - chemistry; Potassium Channels - genetics; Potassium Channels - metabolism; Protein-Serine-Threonine Kinases - metabolism; Sequence Alignment; Shaker Superfamily of Potassium Channels - metabolism; Stress, Physiological - drug effects
• ISSN: 0032-0889; 1532-2548
• DOI: 10.1104/pp.15.01493

In Arabidopsis (Arabidopsis thaliana), the Shaker K⁺ channel AKT1 conducts K⁺ uptake in root cells, and its activity is regulated by CBL1/9-CIPK23 complexes as well as by the AtKC1 channel subunit. CIPK23 and AtKC1 are both involved in the AKT1-mediated low-K⁺ (LK) response; however, the relationship between them remains unclear. In this study, we screened suppressors of low-K⁺ sensitive [lks1 (cipk23)] and isolated the suppressor of lks1 (sls1) mutant, which suppressed the leaf chlorosis phenotype of lks1 under LK conditions. Map-based cloning revealed a point mutation in AtKC1 of sls1 that led to an amino acid substitution (G322D) in the S6 region of AtKC1. The G322D substitution generated a gain-of-function mutation, AtKC1D, that enhanced K⁺ uptake capacity and LK tolerance in Arabidopsis. Structural prediction suggested that glycine-322 is highly conserved in K⁺ channels and may function as the gating hinge of plant Shaker K⁺ channels. Electrophysiological analyses revealed that, compared with wild-type AtKC1, AtKC1D showed enhanced inhibition of AKT1 activity and strongly reduced K⁺ leakage through AKT1 under LK conditions. In addition, phenotype analysis revealed distinct phenotypes of lks1 and atkc1 mutants in different LK assays, but the lks1 atkc1 double mutant always showed a LK-sensitive phenotype similar to that of akt1. This study revealed a link between CIPK-mediated activation and AtKC1-mediated modification in AKT1 regulation. CIPK23 and AtKC1 exhibit distinct effects; however, they act synergistically and balance K⁺ uptake/leakage to modulate AKT1-mediated LK responses in Arabidopsis.