Functional analysis of a calcium-binding transcription factor involved in plant salt stress signaling

Abstract 

Calcium is known to serve as a secondary messenger to mediate salt stress signaling pathway. We found a calcium-binding basic/helix-loop-helix-type transcription factor (AtNIG1) as a salt stress-responsive gene by using the suppression subtractive hybridization. The AtNIG1 was targeted into nucleus and bound to ⁴⁵Ca²⁺, suggesting that AtNIG1 is a nuclear calcium-binding protein. In addition, AtNIG1 bound specifically to the E-box-DNA sequence (CANNTG), which is found in the promoter regions of many salt stress-related genes. Functional analyses with an atnig1-1 knockout mutant revealed that the mutant plants show enhanced sensitivity to salt stress. Further analyses indicated that the atnig1-1 plants have reduced survival rate, fresh weight, chlorophyll content, and protein content upon salt stress, suggesting that the AtNIG1 plays a critical role in plant salt stress signaling. Therefore, this study represents that AtNIG1 is the first known calcium-binding transcription factor involved in plant salt stress signaling.

Abbreviations: bHLH, basic helix-loop-helix, RT, reverse transcription, ABA, abscisic acid, SSH, suppression subtractive hybridization, EMSA, electrophoretic mobility shift assay

Keywords: bHLH, Calcium, Salt stress, Transcription factor, Arabidopsis