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| Motif Analysis and Subcellular Localization of OsUF in Oryza sativa |
| DING Zuo-Mei, WANG Yuan-Yuan, SHI Chao-Nan, WANG Hong, ZHANG Cheng-Long, ZHENG Xiao-Yuan, ZHANG Chao, WU Zu-Jian, WU Jian-Guo* |
| Fujian Province Key Laboratory of Plant Virology/Institute of Plant Virology/Vector-borne Virus Research Center, Fujian Agriculture and Forestry University, Fuzhou 350002, China |
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Abstract E3 ubiquitin ligases, a set of crucial component of ubiquitination pathway, are involved in many aspects of biological process. In order to seek more genes which have key function in antiviral response, this study based on the previous research achievements amplified the Oryza sativa ubiquitin fusion protein (OsUF) cDNA sequence (GenBank No. XM_015757177.2) by using reverse transcription-PCR (RT-PCR) method. The bioinformatics analysis showed that OsUF contained 5 ubiquitin motifs (Ubiquitin, Rad60-SLD, Ubiquitin-2, Rad60-SLD-2 and DUF2407), and 2 non-ubiquitin motif (Ribosomal_L4F01020 and zf-C4pol). Then the full-length of OsUF cDNA was reconstituted into pEarleyGate103, a transient expression vector fused expression with green fluorescent protein (GFP). Agrobacterium-mediated transient expression of OsUF was performed in the epidermal cells of Nicotiana benthamina leaves. Subcellular localization and 4',6-diamidino-2-phenylindole (DAPI) dyeing showed that OsUF protein was localized in the nucleus. Western blotting showed that the GFP-fused OsUF protein was expressed. This analysis of motif and subcellular localization pattern of OsUF may provide key insights into the elucidation of biological function of OsUF and gives more information to learn the mechanism of antiviral response in rice.
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Received: 20 August 2018
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Corresponding Authors:
*, wujianguo81@126.com
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[1] 金伟军, 姚祥春, 吕美巧, 等. 2008. 泛素-蛋白酶体系统的结构、作用和调控机制[J]. 科技通报, 24(1): 29-34.
(Jin W J, Yao X C, Lv M Q, et al.2008. Structure, effect and regulation of the Ubiquitin-proteasome system[J]. Bulletin Science and Technology, 24(1): 29-34.)
[2] Berndsen C E, Wolberger C.2014. New insights into ubiquitin E3 ligase mechanism[J]. Nature Structural and Molecular Biology, 21(4): 301.
[3] Boggio R, Chiocca S.2006. Viruses and sumoylation: Recent highlights[J]. Current Opinion in Microbiology, 9(4): 430-436.
[4] Chao Q, Rothenberg M, Solano R, et al.1997. Activation of the ethylene gas response pathway in Arabidopsis by the nuclear protein ETHYLENE-INSENSITIVE3 and related proteins[J]. Cell, 89(7): 1133-1144.
[5] Chau V, Tobias J W, Bachmair A, et al.1989. A multiubiquitin chain is confined to specific lysine in a targeted short-lived protein[J]. Science, 243(4898): 1576-1584.
[6] Chiu M W, Shih H M, Yang T H, et al.2007. The type 2 dengue virus envelope protein interacts with small ubiquitin-like modifier-1 (SUMO-1) conjugating enzyme 9 (Ubc9)[J]. Journal of Biomedical Science, 14(3): 429-444.
[7] Dong C H, Agarwal M, Zhang Y, et al.2006. The negative regulator of plant cold responses, HOS1, is a RING E3 ligase that mediates the ubiquitination and degradation of ICE1[J]. Proceedings of the National Academy of Sciences, 103(21): 8281-8286.
[8] Guo Q F, Zhang J, Gao Q, et al.2008. Drought tolerance through overexpression of monoubiquitin in transgenic tobacco[J]. Journal of Plant Physiology, 165(16): 1745-1755.
[9] Jia Q, Liu N, Xie K, et al.2016. CLCuMuB βC1 subverts ubiquitination by interacting with NbSKP1s to enhance geminivirus infection in Nicotiana benthamiana[J]. PLoS Pathogens, 12(6): e1005668.
[10] Komander D, Rape M.2012. The ubiquitin code[J]. Annual Review of Biochemistry, 81: 203-229.
[11] Mathieu J, Schwizer S, Martin G B.2014. Pto kinase binds two domains of AvrPtoB and its proximity to the effector E3 ligase determines if it evades degradation and activates plant immunity[J]. PLoS Pathogens,10(7): e1004227.
[12] Metzger M B, Hristova V A, Weissman A M.2012. HECT and RING finger families of E3 ubiquitin ligases at a glance[J]. Journal of Cell Science, 125(3): 531-537.
[13] Meyerson N R, Zhou L G, Guo Y R, et al.2017. Nuclear TRIM25 specifically targets influenza virus ribonucleoproteins to block the onset of RNA chain elongation[J]. Cell Host & Microbe, 22(5): 627-638.
[14] Molinier J, Lechner E, Dumbliauskas E, et al.2008. Regulation and role of Arabidopsis CUL4-DDB1A-DDB2 in maintaining genome integrity upon UV stress[J]. PLoS Genetics, 4(6): e1000093.
[15] Morishita T, Tsutsui Y, Iwasaki H, et al.2002. The Schizosaccharomyces pombe rad60 gene is essential for repairing double-strand DNA breaks spontaneously occurring during replication and induced by DNA-damaging agents[J]. Molecular and Cellular Biology, 22(10): 3537-3548.
[16] Perea-Resa C, Rodríguez-Milla M A, Iniesto E, et al.2017. Prefoldins negatively regulate cold acclimation in Arabidopsis thaliana by promoting nuclear proteasome-mediated HY5 degradation[J]. Molecular Plant, 10(6): 791-804.
[17] Prudden J, Perry J J P, Nie M, et al.2011. DNA repair and global sumoylation are regulated by distinct Ubc9 non-covalent complexes[J]. Molecular and Cellular Biology, 31(11): 2299-2310.
[18] Raab S, Drechsel G, Zarepour Met al.2009. Identification of a novel E3 ubiquitin ligase that is required for suppression of premature senescence in Arabidopsis[J]. The Plant Journal, 59(1): 39-51.
[19] Shen Q, Hu T, Bao M, et al.2016.Tobacco RING E3 ligase NtRFP1 mediates ubiquitination and proteasomal degradation of a geminivirus-encoded βC1[J]. Molecular Plant, 9(6): 911-925.
[20] Smalle J, Vierstra R D.2004.The ubiquitin 26S proteasome proteolytic pathway[J]. Annual Review of Plant Biology, 55: 555-590.
[21] Wang P, Zhao W, Zhao K, et al.2015. TRIM26 negatively regulates interferon-β production and antiviral response through polyubiquitination and degradation of nuclear IRF3[J]. PLoS Pathogens, 11(3): e1004726.
[22] Wu Y C, Deyrieux A F, Wilson V G.2007. Papillomaviruses and the host SUMOylation system[J]. Biochemical Society Transactions, 35(6): 1433-1435.
[23] Zheng N, Shabek N.2017. Ubiquitin ligases: Structure, function, and regulation[J]. Annual Review of Biochemistry, 86: 129-157. |
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