Abstract
The COP9 signalosome (CSN) is a conserved protein complex found in higher eukaryotes, consisting of eight subunits, and it plays a crucial role in regulating various processes of plant growth and development. Among these subunits, CSN2 is one of the most conserved components within the COP9 signalosome complex. Despite its prior identification in other species, its specific function in Oryza sativa L. (Rice) has remained poorly understood. In this study, we investigated the role of CSN2 in rice using gene editing CRISPR/Cas9 technology and overexpression techniques. We created two types of mutants: the oscsn2 mutant and the OsCSN2-OE mutant, both in the background of rice, and also generated point mutants of OsCSN2 (OsCSN2K64E, OsCSN2K67E, OsCSN2K71E and OsCSN2K104E) to further explore the regulatory function of OsCSN2. Phenotypic observation and gene expression analysis were conducted on plants from the generated mutants, tracking their growth from the seedling to the heading stages. The results showed that the loss and modification of OsCSN2 had limited effects on plant growth and development during the early stages of both the wild-type and mutant plants. However, as the plants grew to 60 days, significant differences emerged. The OsCSN2 point mutants exhibited increased tillering compared to the OsCSN2-OE mutant plants, which were already at the tillering stage. On the other hand, the OsCSN2 point mutant had already progressed to the heading and flowering stages, with the shorter plants. These results, along with functional predictions of the OsCSN2 protein, indicated that changes in the 64th, 67th, 71st, and 104th amino acids of OsCSN2 affected its ubiquitination site, influencing the ubiquitination function of CSN and consequently impacting the degradation of the DELLA protein SLR1. Taken together, it can be speculated that OsCSN2 plays a key role in GA and BR pathways by influencing the functional regulation of the transcription factor SLR1 in CSN, thereby affecting the growth and development of rice and the number of tillers.
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References
Achard P, Genschik P (2009) Releasing the brakes of plant growth: how GAs shutdown DELLA proteins. J Exp Bot 60:1085–1092. https://doi.org/10.1093/jxb/ern301
Bajguz A, Tretyn A (2003) The chemical characteristic and distribution of brassinosteroids in plants. Phytochemistry 62:1027–1046. https://doi.org/10.1016/S0031-9422(02)00656-8
Dai C, Xue HW (2010) Rice early flowering1, a CKI, phosphorylates DELLA protein SLR1 to negatively regulate gibberellin signalling. EMBO J 29:1916–1927. https://doi.org/10.1038/emboj.2010.75
Das S, Garhwal V, Gangappa SN (2021) DET1 regulates HY5 through COP1: a new paradigm in the regulation of HY5. Mol Plant 14:864–866. https://doi.org/10.1016/j.molp.2021.05.023
Davière JM, Achard P (2013) Gibberellin signaling in plants. Development 140(1147–1151):9780128096338
Deng XW, Dubiel W, Wei N, Hofmann K, Mundt K, Colicelli J, Kato J, Naumann M, Segal D, Seeger M, Carr A, Glickman M, Chamovitz DA (2000) Unified nomenclature for the COP9 signalosome and its subunits: an essential regulator of development. Trends Genet 16:202–203. https://doi.org/10.1016/S0168-9525(00)01982-X
Dill A, Jung HS, Sun TP (2001) The DELLA motif is essential for gibberellin-induced degradation of RGA. Proc Natl Acad Sci U S A 98:14162–14167. https://www.pnas.org/doi/10.1073/pnas.251534098
Dressel U, Thormeyer D, Altincicek B, Paululat A, Eggert M, Schneider S, Tenbaum SP, Renkawitz R, Baniahmad A (1999) Alien, a highly conserved protein with characteristics of a corepressor for members of the nuclear hormone receptor superfamily. Mol Cell Biol 19:3383–3394. https://doi.org/10.1128/MCB.19.5.3383
Duan E, Wang Y, Li X, Lin Q, Zhang T, Wang Y, Zhou C, Zhang H, Jiang L, Wang J, Lei C, Zhang X, Guo X, Wang H, Wan J (2019) OsSHI1 regulates plant architecture through modulating the transcriptional activity of IPA1 in Rice. Plant Cell 31:1026–1042. https://doi.org/10.1105/tpc.19.00023
Enchev RI, Schreiber A, Beuron F, Morris EP (2010) Structural insights into the COP9 signalosome and its common architecture with the 26S proteasome lid and eIF3. Structure 18:518–527. https://doi.org/10.1016/j.str.2010.02.008
Escher N, Kob R, Tenbaum SP, Eisold M, Baniahmad A, Von Eggeling F, Melle C (2007) Various members of the E2F transcription factor family interact in vivo with the corepressor alien. J Proteome Res 6:1158–1164. https://doi.org/10.1021/pr060500c
Feng S, Martinez C, Gusmaroli G, Wang Y, Zhou J, Wang F, Chen L, Yu L et al (2008) Coordinated regulation of Arabidopsis thaliana development by light and gibberellins. Nature 451:475–479. https://doi.org/10.1038/nature06448
Gao J, Chen H, Yang H, He Y, Tian Z, Li J (2018) A brassinosteroid responsive miRNA-target module regulates gibberellin biosynthesis and plant development. New Phytol 220:488–501. https://doi.org/10.1111/nph.15331
Gomi K, Sasaki A, Itoh H, Ueguchi-Tanaka M, Ashikari M, Kitano H, Matsuoka M (2004) GID2, an F-box subunit of the SCF E3 complex, specifically interacts with phosphorylated SLR1 protein and regulates the gibberellin-dependent degradation of SLR1 in rice. Plant J 37:626–634. https://doi.org/10.1111/j.1365-313x.2003.01990.x
Gusmaroli G, Figueroa P, Serino G, Deng XW (2007) Role of the MPN subunits in COP9 signalosome assembly and activity, and their regulatory interaction with Arabidopsis Cullin3-based E3 ligases. Plant Cell 19:564–581. https://doi.org/10.1105/tpc.106.047571
Han X, Huang X, Deng XW (2020) The photomorphogenic central repressor COP1: conservation and functional diversification during evolution. Plant Commun 1:100044. https://doi.org/10.1016/j.xplc.2020.100044
Harari-Steinberg O, Chamovitz DA (2004) The COP9 signalosome: mediating between kinase signaling and protein degradation. Curr Protein Pept Sci 5:185–189. https://doi.org/10.2174/1389203043379792
He JX, Gendron JM, Sun Y, Gampala SS, Gendron N, Sun CQ, Wang ZY (2005) BZR1 is a transcriptional repressor with dual roles in brassinosteroid homeostasis and growth responses. Science 307:1634–1638. https://doi.org/10.1126/science.1107580
Hofmannv K, Bucher P (1998) The PCI domain: a common theme in three multiprotein complexes. Trends Biochem Sci 23:204–205. https://doi.org/10.1016/S0968-0004(98)01217-1
Ikeda A, Ueguchi-Tanaka M, Sonoda Y, Kitano H, Koshioka M, Futsuhara Y, Matsuoka M, Yamaguchi J (2001) slender rice, a constitutive gibberellin response mutant, is caused by a null mutation of the SLR1 gene, an ortholog of the height-regulating gene GAI/RGA/RHT/D8. Plant Cell 13:999–1010. https://doi.org/10.1105/tpc.13.5.999
Leal JF, Fominaya J, Cascón A, Guijarro MV, Blanco-Aparicio C, Lleonart M, Castro ME, Ramon Y, Cajal S, Robledo M, Beach DH, Carnero A (2007) Cellular senescence bypass screen identifies new putative tumor suppressor genes. Oncogene 27:1961–1970. https://doi.org/10.1038/sj.onc.1210846
Li X, Qian Q, Fu Z, Wang Y, Xiong G, Zeng D, Wang X, Liu X, Teng S, Hiroshi F, Yuan M, Luo D, Han B, Li J (2003) Control of tillering in rice. Nature 422:618–621. https://doi.org/10.1038/NATURE01518
Liao Z, Yu H, Duan J, Yuan K, Yu C, Meng X, Kou L, Chen M, Jing Y, Liu G, Smith SM, Li J (2019) SLR1 inhibits MOC1 degradation to coordinate tiller number and plant height in rice. Nat Commun 10:2738. https://doi.org/10.1038/s41467-019-10667-2
Lier S, Paululat A (2002) The proteasome regulatory particle subunit Rpn6 is required for Drosophila development and interacts physically with signalosome subunit Alien/CSN2. Gene 298:109–119. https://doi.org/10.1016/S0378-1119(02)00930-7
Liu W, Zhang B, He W, Wang Z, Li G, Liu J (2016a) Characterization of in vivo phosphorylation modification of differentially accumulated proteins in cotton fiber-initiation process. Acta Biochim Biophys Sin (shanghai) 48:756–761. https://doi.org/10.1093/abbs/gmw055
Liu Y, Han N, Li Q, Li Z (2016b) Regulatory mechanisms underlying sepsis progression in patients with tumor necrosis factor-α genetic variations. Exp Ther Med 12:323–328. https://doi.org/10.3892/etm.2016.3308
Lyapina S, Cope G, Shevchenko A, Serino G, Tsuge T, Zhou C, Wolf DA, Wei N, Shevchenko A, Deshaies RJ (2001) Promotion of NEDD8-CUL1 conjugate cleavage by COP9 signalosome. Science 292:1382–1385. https://doi.org/10.1126/science.1059780
Millar AH, Heazlewood JL, Giglione C, Holdsworth MJ, Bachmair A, Schulze WX (2019) The scope, functions, and dynamics of posttranslational protein modifications. Annu Rev Plant Biol 70:119–151. https://doi.org/10.1146/annurev-arplant-050718-100211
Musazade E, Liu YX, Ren YX, Wu M, Zeng H, Han SN, Gao XW, Chen SH, Guo LQ (2022) OsCSN1 regulates the growth and development of rice seedlings through the degradation of SLR1 in the GA signaling pathway. Agronomy 12:2946. https://doi.org/10.3390/agronomy12122946
Papaioannou M, Melle C, Baniahmad A (2007) The coregulator Alien. Nucl Recept Signal 5:e008. https://doi.org/10.1621/nrs.05008
Pearce C, Hayden RE, Bunce CM, Khanim FL (2009) Analysis of the role of COP9 Signalosome (CSN) subunits in K562; the first link between CSN and autophagy. BMC Mol Cell Biol 10:31. https://doi.org/10.1186/1471-2121-10-31
Qin N, Xu D, Li J, Deng XW (2020) COP9 signalosome: discovery, conservation, activity, and function. J Integr Plant Biol 62:90–103. https://doi.org/10.1111/jipb.12903
Sasaki A, Itoh H, Gomi K, Ueguchi-Tanaka M, Ishiyama K, Kobayashi M, Jeong DH, An G, Kitano H, Ashikari M, Matsuoka M (2003) Accumulation of phosphorylated repressor for gibberellin signaling in an F-box mutant. Science 299:1896–1898. https://doi.org/10.1126/science.1081077
Scheel H, Hofmann K (2005) Prediction of a common structural scaffold for proteasome lid, COP9-signalosome and eIF3 complexes. BMC Bioinform 6:71. https://doi.org/10.1186/1471-2105-6-71
Schoen A, Yadav I, Wu S, Poland J, Rawat N, Tiwari V (2023) Identification and high-resolution mapping of a novel tiller number gene (tin6) by combining forward genetics screen and MutMap approach in bread wheat. Funct Integr Genomics 23:157. https://doi.org/10.1007/s10142-023-01084-2
Schumacher K, Schmitt T, Rossberg M, Schmitz G, Theres K (1999) The Lateral suppressor (Ls) gene of tomato encodes a new member of the VHIID protein family. Proc Natl Acad Sci USA 96:290–295. https://doi.org/10.1073/pnas.96.1.290
Seeger M, Kraft R, Ferrell K, Bech-Otschir D, Dumdey R, Schade R, Gordon C, Naumann M, Dubiel W (1998) A novel protein complex involved in signal transduction possessing similarities to 26S proteasome subunits. FASEB J 12:469–478. https://doi.org/10.1096/fasebj.12.6.469
Serino G, Deng XW (2003) The COP9 signalosome: regulating plant development through the control of proteolysis. Annu Rev Plant Biol 54:165–182. https://doi.org/10.1146/annurev.arplant.54.031902.134847
Shao GN, Lu ZF, Xiong JS, Wang B, Jing YH, Meng XB, Liu GF, Ma HY, Liang Y, Chen F, Wang YH, Li JY, Yu H (2019) Tiller Bud Formation Regulators MOC1 and MOC3 cooperatively promote tiller bud outgrowth by activating FON1 expression in rice. Mol Plant 12:1090–1102. https://doi.org/10.1016/j.molp.2019.04.008
Sharma A, Ramakrishnan M, Khanna K, Landi M, Prasad R, Bhardwaj R, Zheng B (2022) Brassinosteroids and metalloids: regulation of plant biology. J Hazard Mater 424(Pt C):127518. https://doi.org/10.1016/j.jhazmat.2021.127518
Shi B, Hou J, Yang J, Han IJ, Tu D, Ye S, Yu J, Li L (2023) Genome-wide analysis of the CSN genes in land plants and their expression under various abiotic stress and phytohormone conditions in rice. Gene 850:146905. https://doi.org/10.1016/j.gene.2022.146905
Tong H, Xiao Y, Liu D, Gao S, Liu L, Yin Y, Jin Y, Qian Q, Chu C (2014) Brassinosteroid regulates cell elongation by modulating gibberellin metabolism in rice. Plant Cell 26:4376–4393. https://onlinelibrary.wiley.com/doi/https://doi.org/10.1111/tpj.14692
Tsuji H, Ueguchi-Tanaka M, Nakajima M, Ashikari M, Kitano H, Yamaguchi I, Matsuoka M (2006) Interaction among SLRI1, GID1, and GID2 in the gibberellin signaling pathway in rice cells. Plant Cell Physiol 47:S125. https://doi.org/10.1038/s41598-017-11859-w
Tuller T, Diament A, Yahalom A, Zemach A, Atar S, Chamovitz DA (2019) The COP9 signalosome influences the epigenetic landscape of Arabidopsis thaliana. Bioinformatics 35:2718–2723. https://doi.org/10.1093/bioinformatics/bty1053/5265328
Unterholzner SJ, Rozhon W, Papacek M, Ciomas J, Lange T, Kugler KG, Mayer KF, Sieberer T, Poppenberger B (2015) Brassinosteroids are master regulators of gibberellin biosynthesis in Arabidopsis. Plant Cell 27:2261–2272. https://doi.org/10.1105/tpc.15.00433
Wang D, Musazade E, Wang H, Liu J, Zhang C, Liu W, Liu Y, Guo L (2022) Regulatory mechanism of the constitutive photomorphogenesis 9 signalosome complex in response to abiotic stress in plants. J Agric Food Chem 70:2777–2788. https://doi.org/10.1021/acs.jafc.1c07224
Wang X, Li W, Piqueras R, Cao K, Deng XW, Wei N (2009) Regulation of COP1 nuclear localization by the COP9 signalosome via direct interaction with CSN1. Plant J 58:655–667. https://doi.org/10.1111/j.1365-313x.2009.03805.x
Wang ZY, Bai MY, Oh E, Zhu JY (2012) Brassinosteroid signaling network and regulation of photomorphogenesis. Annu Rev Genet 46:701–724. https://doi.org/10.1146/annurev-genet-102209-163450
Wei N, Deng XW (2003) The COP9 signalosome. Annu Rev Cell Dev Biol 19:261–286. https://doi.org/10.1146/annurev.cellbio.19.111301.112449
Wei N, Serino G, Deng XW (2008) The COP9 signalosome: more than a protease. Trends Biochem Sci 33:592–600. https://doi.org/10.1016/j.tibs.2008.09.004
Xiao Y, Liu D, Zhang G, Tong H, Chu C (2017) Brassinosteroids regulate OFP1, a DLT interacting protein, to modulate plant architecture and grain morphology in rice. Front Plant Sci 8:1698. https://doi.org/10.3389/fpls.2017.01698
Yamaguchi S (2008) Gibberellin metabolism and its regulation. Annu Rev Plant Bio 59:225–251. https://doi.org/10.1007/s003440010039
Ye H, Han GM, Ma Q, Tan YQ, Jiang HY, Zhu SW, Cheng BJ (2015) Effect of temperature on endogenous hormone levels and opposite phyllotaxy in maize leaf primordial. Genet Mol Res 14:17019–17027. https://doi.org/10.4238/2015.December.16.2
Yu JJ, Cui J, Huang H, Cen DC, Liu F, Xu ZF, Wang Y (2024) Identification of flowering genes in Camellia perpetua by comparative transcriptome analysis. Funct Integr Genomics 24:2. https://doi.org/10.1007/s10142-023-01267-x
Zhang HY, Lei G, Zhou HW, He C, Liao JL, Huang YJ (2017) Quantitative iTRAQ-based proteomic analysis of rice grains to assess high night temperature stress. Proteomics 17:1600365. https://doi.org/10.1002/pmic.201600365
Zhang Y, Zeng L (2020) Crosstalk between ubiquitination and other post-translational protein modifications in plant immunity. Plant Commun 1:100041. https://doi.org/10.1016/j.xplc.2020.100041
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This study was funded by the Department of Jilin Province Science & Technology [grant numbers 20230402020GH, 20220402060GH, 20210203011SF].
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H.S.N., W.M., G.L.Q. conceived and designed the experiments. H.S.N., J.T.T., A.B. performed the experiments. H.S.N., Y.W.J., A.B., L.Y.X., W.M., S.K. analyzed and discussed the data, H.S.N., W.M., G.L.Q. wrote and revised the article. All authors contributed to the article and approved the submitted version.
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Han, S., Yue, W., Bao, A. et al. OsCSN2 orchestrates Oryza sativa L. growth and development through modulation of the GA and BR pathways. Funct Integr Genomics 24, 39 (2024). https://doi.org/10.1007/s10142-024-01320-3
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DOI: https://doi.org/10.1007/s10142-024-01320-3