Abstract
E3 ubiquitin ligases catalyze the ubiquitination of a variety of biologically significant protein substrates for targeted degradation through the 26S proteasome, as well as for nonproteolytic regulation of their functions or subcellular localizations. Here we report the identification and characterization of an E3 ubiquitin ligase, the Ring box1 (Rbx1) homologue in maize, which is designated as Zm-Rbx1. Analysis of the genomic organization showed that the gene of Zm-Rbx1 belonged to the chromosome 4 of maize and contained five exons and six introns. Amino acids sequence analysis revealed that Zm-Rbx1 contained conserved cysteine/histidine residues, which are the characteristics of Rbx proteins. Real-time PCR analysis revealed that the expression levels of Zm-Rbx1 increased quickly after salicylic acid, jasmonic acid and sugarcane mosaic virus challenge. Then we suggest that Zm-Rbx1 is involved in the defense response of maize, although detailed molecular mechanism needs to be further studied. After prokaryotic expression and purification of the recombinant Zm-Rbx1 protein from Escherichia coli BL21 (DE3) cells, the ubiquitination assay demonstrated that Zm-Rbx1 showed ubiquitin ligase activity.
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Acknowledgments
This work was supported by the National Natural Science Foundation of China (No. 31101158), Fourth Special Foundation of China Postdoctoral Science Foundation (201104369) and China Postdoctoral Science Foundation (20100470993). We also thank Gabriel Sanglay for critical reading of the manuscript.
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Wu, L., Hu, X., Chen, X. et al. Identification and characterization of an E3 ubiquitin ligase Rbx1 in maize (Zea mays L.). Plant Cell Tiss Organ Cult 116, 253–260 (2014). https://doi.org/10.1007/s11240-013-0391-2
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DOI: https://doi.org/10.1007/s11240-013-0391-2