橡胶树COP9家族基因的克隆及表达分析
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摘要
COP9信号小体(CSN)是进化上保守的蛋白复合体,在茉莉酸信号途径中起着重要的作用。橡胶树的乳管是一种特化的细胞器,是天然橡胶合成和储存的场所。现有的证据表明橡胶的生物合成可能受到茉莉酸信号途径的调控,但是对于茉莉酸信号途径调控天然橡胶的生物合成还了解的不够。本研究采用RACE技术结合RT-PCR从胶乳中克隆了8个CSN基因的全长cDNA序列,根据与拟南芥的相似性,分别命名为HbCSN1～HbCSN8。荧光定量PCR结果显示,8个CSN基因均能在树皮、不同发育时期的叶片、胶乳、雄花和雌花中表达,其中HbCSN5在胶乳中的表达量最高,其他成员在叶片中的表达量最高。而且,部分HbCSNs基因在胶乳中的表达受到割胶和茉莉酸甲酯处理的上调表达。因此,推测这些上调表达的成员可能参与胶乳的茉莉酸信号途径。
The COP9 signalosome(CSN) is an evolutionarily conservative protein complex and plays a crucial role in jasmonate(JA) signaling. The laticifer in rubber tree is specific for rubber biosynthesis. Although it is suggested that rubber biosynthesis in laticifer may be regulated by JA signaling, little is known about the regulatory mechanism of the JA signaling in rubber biosynthesis. In this study, the full-length cDNAs of eight CSN genes(designated as HbCSN1 to HbCSN8) which were respectively related to the eight CSN genes in Arabidopsis were cloned from the laticifer of rubber trees. The differentially expressed pattern of the eight CSN genes among bark tissues, leaves at the different development stages, latex, male and female flowers was revealed by real-time quantitative RT-PCR. The result showed that the eight HbCSNs were differentially expressed in all the tested tissues. Of which, HbCSN5 was the most abundance in latex,and the other HbCSNs were mainly expressed in the leaf. Additionally, most of the eight CSN genes in laticifer were up-regulated by tapping and methyl jasmonate(MeJA). The differentially responsive CSN members indicated that the COP9 signalosome may be involved in the JA signalings of latex in rubber trees.
The COP9 signalosome(CSN) is an evolutionarily conservative protein complex and plays a crucial role in jasmonate(JA) signaling. The laticifer in rubber tree is specific for rubber biosynthesis. Although it is suggested that rubber biosynthesis in laticifer may be regulated by JA signaling, little is known about the regulatory mechanism of the JA signaling in rubber biosynthesis. In this study, the full-length cDNAs of eight CSN genes(designated as HbCSN1 to HbCSN8) which were respectively related to the eight CSN genes in Arabidopsis were cloned from the laticifer of rubber trees. The differentially expressed pattern of the eight CSN genes among bark tissues, leaves at the different development stages, latex, male and female flowers was revealed by real-time quantitative RT-PCR. The result showed that the eight HbCSNs were differentially expressed in all the tested tissues. Of which, HbCSN5 was the most abundance in latex,and the other HbCSNs were mainly expressed in the leaf. Additionally, most of the eight CSN genes in laticifer were up-regulated by tapping and methyl jasmonate(MeJA). The differentially responsive CSN members indicated that the COP9 signalosome may be involved in the JA signalings of latex in rubber trees.
引文
[1]Wei N,Serino G,Deng X W.The COP9 signalosome:more than a protease[J].Trends in Biochemical Sciences,2008,33(12):592-600.
[2]Stratmann J W,Gusmaroli G.Many jobs for one good copthe COP9 signalosome guards development and defense[J].Plant Science,2012,185-186:50-64.
[3]Wei N,Deng X W.COP9:a new genetic locus involved in light-regulated development and gene expression in Arabidopsis[J].Plant Cell,1992,4(12):1507-1518.
[4]Wei N,Chamovitz D A,Deng X W.Arabidopsis COP9 is a component of a novel signaling complex mediating light control of development[J].Cell,1994,78(1):117-124.
[5]Chamovitz D A,Wei N,Osterlund M T,et al.The COP9complex,a novel multisubunit nuclear regulator involved in light control of a plant developmental switch[J].Cell,1996,86(1):115-121.
[6]Wei N,Deng X W.The COP9 signalosome[J].Annual Review of Cell and Developmental Biology,2003,19:261-286.
[7]Hofmann K,Bucher P.The PCI domain:a common theme in three multiprotein complexes[J].Trends in Biochemical Sciences,1998,23(6):204-205.
[8]Cope G A,Suh G S,Aravind L,et al.Role of predicted metalloprotease motif of Jab1/Csn5 in cleavage of Nedd8 from Cul1[J].Science,2002,298(5593):608-611.
[9]Lyapina S,Cope G,Shevchenko A,et al.Promotion of NEDD-CUL1 conjugate cleavage by COP9 signalosome[J].Science,2001,292(5520):1382-1385.
[10]Schwechheimer C,Serino G,Callis J,et al.Interactions of the COP9 signalosome with the E3 ubiquitin ligase SCFTIRIin mediating auxin response[J].Science,2001,292(5520):1379-1382.
[11]Groisman R,Polanowska J,Kuraoka I,et al.The ubiquitin ligase activity in the DDB2 and CSA complexes is differentially regulated by the COP9 signalosome in response to DNA damage[J].Cell,2003,113(3):357-367.
[12]Pintard L,Kurz T,Glaser S,et al.Neddylation and deneddylation of CUL-3 is required to target MEI-1/Katanin for degradation at the meiosis-to-mitosis transition in C.elegans[J].Current Biology,2003,13(11):911-921.
[13]Stuttmann J,Lechner E,Guerois R,et al.COP9 signalosome-and 26S proteasome-dependent regulation of SCFTIR1accumulation in Arabidopsis[J].Journal of Biological Chemistry,2009,284(12):7920-7930.
[14]Wang X,Feng S,Nakayama N,et al.The COP9 signalosome interacts with SCFUFO and participates in Arabidopsis flower development[J].Plant Cell,2003,15(5):1071-1082.
[15]Feng S,Ma L,Wang X,et al.The COP9 signalosome interacts physically with SCFCOI1 and modulates jasmonate responses[J].Plant Cell,2003,15(5):1083-1094.
[16]Lozano-Duran R,Rosas-Diaz T,Gusmaroli G,et al.Geminiviruses subvert ubiquitination by altering CSN-mediated derubylation of SCF E3 ligase complexes and inhibit jasmonate signaling in Arabidopsis thaliana[J].Plant Cell,2011,23(3):1014-1032.
[17]Dohmann E M,Nill C,Schwechheimer C.DELLA proteins restrain germination and elongation growth in Arabidopsis thaliana COP9 signalosome mutants[J].European Journal of Cell Biology,2010,89(2-3):163-168.
[18]Jin D,Wu M,Li B,et al.The COP9 Signalosome regulates seed germination by facilitating protein degradation of RGL2and ABI5[J].PLoS Genetics,2018,14(2):e1007237.
[19]Franciosini A,Lombardi B,Iafrate S,et al.The Arabidopsis COP9 SIGNALOSOME INTERACTING F-BOX KELCH 1protein forms an SCF ubiquitin ligase and regulates hypocotyl elongation[J].Molecular Plant,2013,6(5):1616-1629.
[20]Deng X,Guo D,Yang S,et al.Jasmonate signalling in the regulation of rubber biosynthesis in laticifer cells of rubber tree,Hevea brasiliensis[J].Journal of Experimental Botany,2018,69(15):3559-3571.
[21]Hao B Z,Wu J L.Laticifer differentiation in Hevea brasiliensis:induction by exogenous jasmonic acid and linolenic acid[J].Annals of Botany,2000,85:37-43.
[22]Chao J Q,Chen Y Y,Wu S H,et al.Comparative transcriptome analysis of latex from rubber tree clone CATAS8-79and PR107 reveals new cues for the regulation of latex regeneration and duration of latex flow[J].BMC Plant Biology,2015,15:104.doi:10.1186/s12870-015-0488-3.
[23]Hu B,Jin J P,Guo A Y,et al.GSDS 2.0:an upgraded gene feature visualization server[J].Bioinformatics,2015,31(8):1296-1297.
[24]Zhao Y,Zhou L M,Chen Y Y,et al.MYC genes with differential responses to tapping,mechanical wounding,ethephon and methyl jasmonate in laticifers of rubber tree(Hevea brasiliensis Muell.Arg.)[J].Journal of Plant Physiology,2011,168(14):1649-1658.
[25]Wang Y,Guo D,Li H L,et al.Characterization of HbWRKY1,a WRKY transcription factor from Hevea brasiliensis that negatively regulates Hb SRPP[J].Plant Physiology Biochemistry,2013,71:283-289.
[26]Li H L,Guo D,Yang Z P,et al.Genome-wide identification and characterization of WRKY gene family in Hevea brasiliensis[J].Genomics,2014,104(1):14-23.
[27]Kapelari B,Bech-Otschir D,Hegerl R,et al.Electron microscopy and subunit-subunit interaction studies reveal a first architecture of COP9 signalosome[J].Journal of Molecular Biology,2000,300(5):1169-1178.
[28]Tsuge T,Matsui M,Wei N.The subunit 1 of the COP9 signalosome suppresses gene expression through its N-terminal domain and incorporates into the complex through the PCIdomain[J].Journal of Molecular Biology,2001,305(1):1-9.
[29]Nezames C D,Deng X W.The COP9 signalosome:its regulation of cullin-based E3 ubiquitin ligases and role in photomorphogenesis[J].Plant Physiology,2012,160(1):38-46.
[30]Maytal-Kivity V,Reis N,Hofmann K,et al.MPN+,a putative catalytic motif found in a subset of MPN domain proteins from eukaryotes and prokaryotes,is critical for Rpn11function[J].BMC Biochemistry,2002,3:28.
[31]Tran H J,Allen M D,Lowe J,et al.Structure of the Jab1/MPN domain and its implications for proteasome function[J].Biochemistry,2003,42(39):11460-11465.
[32]Ambroggio X I,Rees D C,Deshaies R J.JAMM:a metalloprotease-like zinc site in the proteasome and signalosome[J].PLoS Biology,2004,2(1):E2.
[33]Kotiguda G G,Weinberg D,Dessau M,et al.The organization of a CSN5-containing subcomplex of the COP9 signalosome[J].Journal of Biology Chemistry,2012,287(50):42031-42041.
[34]Jin D,Li B,Deng X W,et al.Plant COP9 signalosome subunit 5,CSN5[J].Plant Science,2014,224:54-61.
[35]Hind S R,Pulliam S E,Veronese P,et al.The COP9 signalosome controls jasmonic acid synthesis and plant responses to herbivory and pathogens[J].Plant Journal,2011,65(3):480-491.
[36]Peng S Q,Xu J,Li H L,et al.Cloning and molecular characterization of Hb COI1 from Hevea brasiliensis[J].Bioscience Biotechnology and Biochemistry,2009,73(3):665-670.
[37]Tian W M,Huang W F,Zhao Y.Cloning and characterization of HbJAZ1 from the laticifer cells in rubber tree(Hevea brasiliensis Muell.Arg.)[J].Trees,2010,24:771-779.
[38]Liu J P,Hu J,Liu Y H,et al.Transcriptome analysis of Hevea brasiliensis in response to exogenous methyl jasmonate provides novel insights into regulation of jasmonate-elicited rubber biosynthesis[J].Physiology and Molecular Biology of Plants,2018,24(3):349-358.
[2]Stratmann J W,Gusmaroli G.Many jobs for one good copthe COP9 signalosome guards development and defense[J].Plant Science,2012,185-186:50-64.
[3]Wei N,Deng X W.COP9:a new genetic locus involved in light-regulated development and gene expression in Arabidopsis[J].Plant Cell,1992,4(12):1507-1518.
[4]Wei N,Chamovitz D A,Deng X W.Arabidopsis COP9 is a component of a novel signaling complex mediating light control of development[J].Cell,1994,78(1):117-124.
[5]Chamovitz D A,Wei N,Osterlund M T,et al.The COP9complex,a novel multisubunit nuclear regulator involved in light control of a plant developmental switch[J].Cell,1996,86(1):115-121.
[6]Wei N,Deng X W.The COP9 signalosome[J].Annual Review of Cell and Developmental Biology,2003,19:261-286.
[7]Hofmann K,Bucher P.The PCI domain:a common theme in three multiprotein complexes[J].Trends in Biochemical Sciences,1998,23(6):204-205.
[8]Cope G A,Suh G S,Aravind L,et al.Role of predicted metalloprotease motif of Jab1/Csn5 in cleavage of Nedd8 from Cul1[J].Science,2002,298(5593):608-611.
[9]Lyapina S,Cope G,Shevchenko A,et al.Promotion of NEDD-CUL1 conjugate cleavage by COP9 signalosome[J].Science,2001,292(5520):1382-1385.
[10]Schwechheimer C,Serino G,Callis J,et al.Interactions of the COP9 signalosome with the E3 ubiquitin ligase SCFTIRIin mediating auxin response[J].Science,2001,292(5520):1379-1382.
[11]Groisman R,Polanowska J,Kuraoka I,et al.The ubiquitin ligase activity in the DDB2 and CSA complexes is differentially regulated by the COP9 signalosome in response to DNA damage[J].Cell,2003,113(3):357-367.
[12]Pintard L,Kurz T,Glaser S,et al.Neddylation and deneddylation of CUL-3 is required to target MEI-1/Katanin for degradation at the meiosis-to-mitosis transition in C.elegans[J].Current Biology,2003,13(11):911-921.
[13]Stuttmann J,Lechner E,Guerois R,et al.COP9 signalosome-and 26S proteasome-dependent regulation of SCFTIR1accumulation in Arabidopsis[J].Journal of Biological Chemistry,2009,284(12):7920-7930.
[14]Wang X,Feng S,Nakayama N,et al.The COP9 signalosome interacts with SCFUFO and participates in Arabidopsis flower development[J].Plant Cell,2003,15(5):1071-1082.
[15]Feng S,Ma L,Wang X,et al.The COP9 signalosome interacts physically with SCFCOI1 and modulates jasmonate responses[J].Plant Cell,2003,15(5):1083-1094.
[16]Lozano-Duran R,Rosas-Diaz T,Gusmaroli G,et al.Geminiviruses subvert ubiquitination by altering CSN-mediated derubylation of SCF E3 ligase complexes and inhibit jasmonate signaling in Arabidopsis thaliana[J].Plant Cell,2011,23(3):1014-1032.
[17]Dohmann E M,Nill C,Schwechheimer C.DELLA proteins restrain germination and elongation growth in Arabidopsis thaliana COP9 signalosome mutants[J].European Journal of Cell Biology,2010,89(2-3):163-168.
[18]Jin D,Wu M,Li B,et al.The COP9 Signalosome regulates seed germination by facilitating protein degradation of RGL2and ABI5[J].PLoS Genetics,2018,14(2):e1007237.
[19]Franciosini A,Lombardi B,Iafrate S,et al.The Arabidopsis COP9 SIGNALOSOME INTERACTING F-BOX KELCH 1protein forms an SCF ubiquitin ligase and regulates hypocotyl elongation[J].Molecular Plant,2013,6(5):1616-1629.
[20]Deng X,Guo D,Yang S,et al.Jasmonate signalling in the regulation of rubber biosynthesis in laticifer cells of rubber tree,Hevea brasiliensis[J].Journal of Experimental Botany,2018,69(15):3559-3571.
[21]Hao B Z,Wu J L.Laticifer differentiation in Hevea brasiliensis:induction by exogenous jasmonic acid and linolenic acid[J].Annals of Botany,2000,85:37-43.
[22]Chao J Q,Chen Y Y,Wu S H,et al.Comparative transcriptome analysis of latex from rubber tree clone CATAS8-79and PR107 reveals new cues for the regulation of latex regeneration and duration of latex flow[J].BMC Plant Biology,2015,15:104.doi:10.1186/s12870-015-0488-3.
[23]Hu B,Jin J P,Guo A Y,et al.GSDS 2.0:an upgraded gene feature visualization server[J].Bioinformatics,2015,31(8):1296-1297.
[24]Zhao Y,Zhou L M,Chen Y Y,et al.MYC genes with differential responses to tapping,mechanical wounding,ethephon and methyl jasmonate in laticifers of rubber tree(Hevea brasiliensis Muell.Arg.)[J].Journal of Plant Physiology,2011,168(14):1649-1658.
[25]Wang Y,Guo D,Li H L,et al.Characterization of HbWRKY1,a WRKY transcription factor from Hevea brasiliensis that negatively regulates Hb SRPP[J].Plant Physiology Biochemistry,2013,71:283-289.
[26]Li H L,Guo D,Yang Z P,et al.Genome-wide identification and characterization of WRKY gene family in Hevea brasiliensis[J].Genomics,2014,104(1):14-23.
[27]Kapelari B,Bech-Otschir D,Hegerl R,et al.Electron microscopy and subunit-subunit interaction studies reveal a first architecture of COP9 signalosome[J].Journal of Molecular Biology,2000,300(5):1169-1178.
[28]Tsuge T,Matsui M,Wei N.The subunit 1 of the COP9 signalosome suppresses gene expression through its N-terminal domain and incorporates into the complex through the PCIdomain[J].Journal of Molecular Biology,2001,305(1):1-9.
[29]Nezames C D,Deng X W.The COP9 signalosome:its regulation of cullin-based E3 ubiquitin ligases and role in photomorphogenesis[J].Plant Physiology,2012,160(1):38-46.
[30]Maytal-Kivity V,Reis N,Hofmann K,et al.MPN+,a putative catalytic motif found in a subset of MPN domain proteins from eukaryotes and prokaryotes,is critical for Rpn11function[J].BMC Biochemistry,2002,3:28.
[31]Tran H J,Allen M D,Lowe J,et al.Structure of the Jab1/MPN domain and its implications for proteasome function[J].Biochemistry,2003,42(39):11460-11465.
[32]Ambroggio X I,Rees D C,Deshaies R J.JAMM:a metalloprotease-like zinc site in the proteasome and signalosome[J].PLoS Biology,2004,2(1):E2.
[33]Kotiguda G G,Weinberg D,Dessau M,et al.The organization of a CSN5-containing subcomplex of the COP9 signalosome[J].Journal of Biology Chemistry,2012,287(50):42031-42041.
[34]Jin D,Li B,Deng X W,et al.Plant COP9 signalosome subunit 5,CSN5[J].Plant Science,2014,224:54-61.
[35]Hind S R,Pulliam S E,Veronese P,et al.The COP9 signalosome controls jasmonic acid synthesis and plant responses to herbivory and pathogens[J].Plant Journal,2011,65(3):480-491.
[36]Peng S Q,Xu J,Li H L,et al.Cloning and molecular characterization of Hb COI1 from Hevea brasiliensis[J].Bioscience Biotechnology and Biochemistry,2009,73(3):665-670.
[37]Tian W M,Huang W F,Zhao Y.Cloning and characterization of HbJAZ1 from the laticifer cells in rubber tree(Hevea brasiliensis Muell.Arg.)[J].Trees,2010,24:771-779.
[38]Liu J P,Hu J,Liu Y H,et al.Transcriptome analysis of Hevea brasiliensis in response to exogenous methyl jasmonate provides novel insights into regulation of jasmonate-elicited rubber biosynthesis[J].Physiology and Molecular Biology of Plants,2018,24(3):349-358.