E3 ubiquitin ligases are participated in numerous processes, regulating the response to biotic and abiotic stresses. Botrytis susceptible1 interactor (BOI) is a RING (Really Interesting New Gene)-type E3 ligase that mediates the ubiquitination of BOS1 (Botrytis susceptible1), a transcription factor involved in stress and pathogen responses. Although BOI is an E3 ligase, there are reports to show that BOI interacts with target proteins such as DELLAs or CONSTANS to repress gibberellin responses and flowering without the degradation of the target proteins. In this article, we utilize diversified methods to comprehensively analyze the expression pattern, interaction network and function of BOI gene. Firstly, 1800 bp upstream region of BOI gene from Arabidopsisthaliana (Arabidopsis) genome was isolated, and fused GUS reporter gene. The resulting expression cassette was introduced into wild-type Arabidopsis through Agrobacterium-mediated transformation. The result demonstrated that BOI gene was expressed predominantly in leaves, siliques, young roots, and flowering tissues, indicating that BOI gene may be involved in multiple processes in plant growth and development in Arabidopsis. Besides, eight candidate interacting proteins were obtained from the Arabidopsis cDNA library via yeast two-hybrid technology, including EXO70E2 (AT5G61010), WRKY7 (AT4G24240), WRKY11 (AT4G31550), WRKY17 (AT2G24570), UBP20 (AT4G17895), L5 (AT1G12290), SAUR9 (AT4G36110) and TCP21 (AT5G08330). Functional analysis of these candidate interacting proteins manifested that they related to multiple pathways, including biological and abiotic stress, programmed cell death, protein degradation, material metabolism and transcriptional regulation. In addition, the results of the transient assay proclaimed that BOI protein affects the protein stability of EXO70E2 and L5 through its E3 ubiquitin ligase activity. Our results provide novel clues for a better understanding of molecular mechanisms underlying BOI-mediated regulations.
References
[1]
Rosenthal, M.R. and Ng, C.L. (2021) A Proteasome Mutation Sensitizes P. falciparum Cam3.ii K13c580y Parasites to DHA and OZ439. ACS Infectious Diseases, 7, 1923-1931. https://doi.org/10.1021/acsinfecdis.0c00900
[2]
Wang, B., Hu, H., Wang, X., Shao, Z., Shi, D., Wu, F., et al. (2024) POLE2 Promotes Osteosarcoma Progression by Enhancing the Stability of CD44. Cell Death Discovery, 10, Article No. 177. https://doi.org/10.1038/s41420-024-01875-x
[3]
Tang, W., Cai, D., Fu, Y., Zheng, Z., Huang, X., Khouzam, R.N., et al. (2023) 4-phenylbutyric Acid Re-Trafficking hERG/G572R Channel Protein by Modulating the Endoplasmic Reticulum Stress-Associated Chaperones and Endoplasmic Reticulum-Associated Degradation Gene. Journal of Thoracic Disease, 15, 4472-4485. https://doi.org/10.21037/jtd-23-1252
[4]
Yu, Y., Yang, S., Bian, L., Yu, K., Meng, X., Zhang, G., et al. (2021) Identification of C3H2C3-Type RING E3 Ubiquitin Ligase in Grapevine and Characterization of Drought Resistance Function of VyRCHC114. BMC Plant Biology, 21, Article No. 422. https://doi.org/10.1186/s12870-021-03162-8
[5]
Liang, Y., Wang, Z., Wang, Q., Zhou, X. and Qian, Y. (2023) The Ring-Finger Protein NbRFP1 Contributes to Regulating the Host Hypersensitive Response Induced by Oat Dwarf Virus Repa. International Journal of Molecular Sciences, 24, Article 7697. https://doi.org/10.3390/ijms24097697
[6]
Luo, H., Laluk, K., Lai, Z., Veronese, P., Song, F. and Mengiste, T. (2010) The Arabidopsis Botrytis Susceptible1 Interactor Defines a Subclass of RING E3 Ligases That Regulate Pathogen and Stress Responses. Plant Physiology, 154, 1766-1782. https://doi.org/10.1104/pp.110.163915
[7]
Park, J., Nguyen, K.T., Park, E., Jeon, J. and Choi, G. (2013) DELLA Proteins and Their Interacting RING Finger Proteins Repress Gibberellin Responses by Binding to the Promoters of a Subset of Gibberellin-Responsive Genes in Arabidopsis. The Plant Cell, 25, 927-943. https://doi.org/10.1105/tpc.112.108951
[8]
Nguyen, K.T., Park, J., Park, E., Lee, I. and Choi, G. (2015) The Arabidopsis RING Domain Protein BOI Inhibits Flowering via Co-Dependent and Co-Independent Mechanisms. Molecular Plant, 8, 1725-1736. https://doi.org/10.1016/j.molp.2015.08.005
[9]
Huang, J., Wu, X. and Gao, Z. (2021) The Ring-Type Protein BOI Negatively Regulates the Protein Level of a CC-NBS-LRR in Arabidopsis. Biochemical and Biophysical Research Communications, 578, 104-109. https://doi.org/10.1016/j.bbrc.2021.09.038
[10]
Huang, J., Wu, X. and Gao, Z. (2021) A Nucleocytoplasmic-Localized E3 Ligase Affects the NLR Receptor Stability. Biochemical and Biophysical Research Communications, 583, 1-6. https://doi.org/10.1016/j.bbrc.2021.10.052
[11]
Huang, J., Guan, X., Zhong, X., Jia, P., Zhang, H. and Ruan, H. (2024) Structural and Functional Insights into an Arabidopsis NBS-LRR Receptor in Nicotiana benthamiana. American Journal of Molecular Biology, 14, 84-96. https://doi.org/10.4236/ajmb.2024.142007
[12]
Zhang, H., Zheng, D., Song, F. and Jiang, M. (2022) Expression Patterns and Functional Analysis of 11 E3 Ubiquitin Ligase Genes in Rice. Frontiers in Plant Science, 13, Article 840360. https://doi.org/10.3389/fpls.2022.840360
[13]
Kumar, A., Sichov, N., Bucki, P. and Miyara, S.B. (2023) SLWRKY16 and SLWRKY31 of Tomato, Negative Regulators of Plant Defense, Involved in Susceptibility Activation Following Root-Knot Nematode Meloidogyne javanica Infection. Scientific Reports, 13, Article No. 14592. https://doi.org/10.1038/s41598-023-40557-z
[14]
Chen, H., Lai, Z., Shi, J., Xiao, Y., Chen, Z. and Xu, X. (2010) Roles of Arabidopsis WRKY18, WRKY40 and WRKY60 Transcription Factors in Plant Responses to Abscisic Acid and Abiotic Stress. BMC Plant Biology, 10, Article No. 281. https://doi.org/10.1186/1471-2229-10-281
[15]
Li, Z., Huang, J., Hu, Y., Zhou, X., Tan, X., Wang, Z., et al. (2024) The Ring-Type E3 Ligase BOI Interacts with EXO70E2 and Mediates Its Ubiquitination in Arabidopsis. Life, 14, Article 1169. https://doi.org/10.3390/life14091169
[16]
Hou, H., Fang, J., Liang, J., Diao, Z., Wang, W., Yang, D., et al. (2020) OsEXo70B1 Positively Regulates Disease Resistance to Magnaporthe oryzae in Rice. International Journal of Molecular Sciences, 21, Article 7049. https://doi.org/10.3390/ijms21197049
[17]
De la Concepcion, J.C. (2023) The Exocyst Complex Is an Evolutionary Battleground in Plant-Microbe Interactions. Current Opinion in Plant Biology, 76, Article ID: 102482. https://doi.org/10.1016/j.pbi.2023.102482
[18]
Seo, H.S., Yang, J., Ishikawa, M., Bolle, C., Ballesteros, M.L. and Chua, N. (2003) LAF1 Ubiquitination by COP1 Controls Photomorphogenesis and Is Stimulated by SPA1. Nature, 423, 995-999. https://doi.org/10.1038/nature01696
[19]
Amemiya, Y., Azmi, P. and Seth, A. (2008) Autoubiquitination of BCA2 RING E3 Ligase Regulates Its Own Stability and Affects Cell Migration. Molecular Cancer Research, 6, 1385-1396. https://doi.org/10.1158/1541-7786.mcr-08-0094
[20]
Wu, X., Huang, J., Cao, Y. and Gao, Z. (2021) The Resistance Associated Protein RIN4 Promotes the Extracellular Transport of AtEXO70E2. Biochemical and Biophysical Research Communications, 555, 40-45. https://doi.org/10.1016/j.bbrc.2021.03.072
