Abstract
SUMOylation in plants is associated with biotic and abiotic stress responses, flowering, and other aspects of development. ICE1 protein SUMOylation by the SUMO E3 ligase SIZ1 plays important roles in plant cold tolerance. Here, we reported the subcellular localization of EcaICE1 and its interaction with EcaSIZ1 in Eucalyptus camaldulensis. The genes EcaICE1 and EcaSIZ1 were isolated by homologous cloning. The subcellular localization analysis showed that EcaICE1 was located in nucleus. Bimolecular fluorescence complementation (BiFC) analysis revealed that EcaICE1 could interact with EcaSIZ1 in the nucleus of Nicotiana benthamiana leaves. Moreover, yeast two-hybrid assay confirmed that the amino acid region from position 84 to 126 in EcaICE1 was critical for the strong transactivation activity of EcaICE1 and that the C-terminal region from position 361 to 557 in EcaICE1 was the key region for its interaction with EcaSIZ1 using different truncated lengths of non-transactivation activity of EcaICE1 as the bait protein. Collectively, our results showed that EcaICE1 may have a SUMOylation pathway similar to Arabidopsis thaliana.
Highlights
SUMOylation in plants is associated with biotic and abiotic stress responses, flowering and other aspects of development, and ICE1 protein SUMOylation by SUMO E3 ligase SIZ1 plays important roles in plant cold tolerance
In Arabidopsis, the cold signaling pathways, ICE-CBF-cold-regulated genes (CORs) pathway (Ding et al 2015; Shi et al 2018) and ICE-CBF pathway is positively correlated with response to low temperature stress (Lin et al 2014), had been studied clearly while the research of subcellular localization, transcriptional activation and interactions with upstream regulators in E
This study indicated that 84th to 125th amino acid of N-terminal of protein was the critical region of transcriptional activation in EcaICE1 of E. camaldulensis, including the Q-rich domain as well as according with the transcriptional activation structure of classical transcription factor Sp1
Summary
SUMOylation in plants is associated with biotic and abiotic stress responses, flowering and other aspects of development, and ICE1 protein SUMOylation by SUMO E3 ligase SIZ1 plays important roles in plant cold tolerance. CBFs or DREB1s are key transcription factors of low temperature signaling pathway and play important roles in enhancing plant cold tolerance (Chinnusamy et al 2007). SUMO E3 ligase SIZ1, can enhance the stability of ICE1 protein by SUMOylation and promote the expression of CBF3, and causes enhanced cold tolerance (Miura et al 2007). ICE1 gene had been isolated from A. thaliana (Chinnusamy et al 2003), Brassica juncea (Wang et al 2005), Populus suaveolens (Lin et al.2007), wheat (Badawi et al 2008), banana (ZHAO et al 2013), E. camaldulensis (Lin et al 2014), Pyrus ussuriensis (Huang et al 2015) and Hevea brasiliensis (Deng et al 2017), the understanding of its positive and negative regulations is still limited in woody plants
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