小桐子MAPKKKK基因家族的全基因组鉴定及表达分析
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摘要
促分裂原活化蛋白激酶激酶激酶激酶(mitogen-activated protein kinase kinase kinase kinase, MAPKKKK)属Ser/Thr类蛋白激酶,参与典型MAPK级联系统与其它信号转导途径,从而调节植物光周期、花器官生长及抗逆性等生物学过程。本研究基于序列比对的方法,在全基因组水平对小桐子MAPKKKK基因家族进行鉴定,并对其基因结构、系统进化、表达特性及潜在功能进行了解析。结果表明,小桐子基因组中共鉴定到6个MAPKKKK基因,主要定位细胞核,蛋白序列长度分布在513～812 aa,等电点分布在5.12～6.72。序列比对都发现激酶结构域及保守基序-TFVGTPxWMAPEV-,除JcMAPKKKK4激酶结构域位于序列中部外,其它成员都位于N端。与拟南芥、葡萄MAPKKKK共聚类与系统进化分析显示, MAPKKKK聚类为8个亚组,且各亚组内成员存在蛋白序列长度、外显子数量、等电点特异性。基因结构分析表明,除JcMAPKKKK6仅含有1个外显子,其它5个小桐子MAPKKKK基因包含18～22个外显子。表达分析显示,JcMAPKKKK1、JcMAPKKKK2及JcMAPKKKK5在叶片中表达量较高,而JcMAPKKKK3与JcMAPKKKK6在根中表达量较高。同时,JcMAPKKKK3与JcMAPKKKK6是响应小桐子抗冷性的主要基因。蛋白互作分析表明,小桐子MAPKKKK与WNK、Mo25及Mob家族蛋白具有广泛的互作关系,并可能参与植物极性生长、细胞分裂及ABA信号转导等过程。以上结果为开展小桐子MAPKKKK基因的功能鉴定与信号转导机制研究提供了参考。
Mitogen-activated protein kinase kinase kinase kinase, as a Ser/Thr protein kinase, function as the biological processes of photoperiod, flowering time, and stress resistance, which plays vital roles in classical MAPK cascade and other signalling transduction pathways. For insight into the characteristics of MAPKKKK in Jatropha curcas, the MAPKKKK gene family was identified from J. curcas based on the BLAST method, and then the gene structure, phylogenetic relationship, expression profile and potential function were analyzed. The results showed that the J. curcas genome identified 6 MAPKKKKs and the predicted MAPKKKK proteins mainly located in nucleus with protein length ranged from 513 to 812 aa and pI value distributed from 5.12 to6.72. Kinase domain and conserved motif of-TFVGTPxWMAPEV-were found in all the J. curcas MAPKKKK depends on sequence alignment. Except for JcMAPKKKK4 located in the middle of polypeptide, all other members located at N-terminal. Based on the phylogenetic relationship, MAPKKKKs from J. curcas with Arabidopsis thaliana and Vitis vinifera were classified into 8 subgroups and owned subgroup-specificity in protein length, exon number, and electrical point. Furthermore, gene structure analysis showed that except for only one exon in JcMAPKKKK6, the other five J. curcas MAPKKKK genes contained 18–22 exons. Expression analysis revealed that JcMAPKKKK1, JcMAPKKKK2, and Jc MAPKKKK5 were highly expressed in leaves, but JcMAPKKKK3 and JcMAPKKKK6 possessed high expression levels in roots. In addition, JcMAPKKKK3 and JcMAPKKKK6 were the main genes that respond to the cold resistance in J. curcas. Protein interaction analysis indicated that J. curcas MAPKKKKs had extensive interaction with WNKN, Mo25, and Mob family proteins, and were involved in plant polarity growth, cell division, and ABA signal transduction. The results of this study might lay a significant foundation for further studies on the gene function and signalling transduction mechanism of MAPKKKK gene family in J. curcas.
Mitogen-activated protein kinase kinase kinase kinase, as a Ser/Thr protein kinase, function as the biological processes of photoperiod, flowering time, and stress resistance, which plays vital roles in classical MAPK cascade and other signalling transduction pathways. For insight into the characteristics of MAPKKKK in Jatropha curcas, the MAPKKKK gene family was identified from J. curcas based on the BLAST method, and then the gene structure, phylogenetic relationship, expression profile and potential function were analyzed. The results showed that the J. curcas genome identified 6 MAPKKKKs and the predicted MAPKKKK proteins mainly located in nucleus with protein length ranged from 513 to 812 aa and pI value distributed from 5.12 to6.72. Kinase domain and conserved motif of-TFVGTPxWMAPEV-were found in all the J. curcas MAPKKKK depends on sequence alignment. Except for JcMAPKKKK4 located in the middle of polypeptide, all other members located at N-terminal. Based on the phylogenetic relationship, MAPKKKKs from J. curcas with Arabidopsis thaliana and Vitis vinifera were classified into 8 subgroups and owned subgroup-specificity in protein length, exon number, and electrical point. Furthermore, gene structure analysis showed that except for only one exon in JcMAPKKKK6, the other five J. curcas MAPKKKK genes contained 18–22 exons. Expression analysis revealed that JcMAPKKKK1, JcMAPKKKK2, and Jc MAPKKKK5 were highly expressed in leaves, but JcMAPKKKK3 and JcMAPKKKK6 possessed high expression levels in roots. In addition, JcMAPKKKK3 and JcMAPKKKK6 were the main genes that respond to the cold resistance in J. curcas. Protein interaction analysis indicated that J. curcas MAPKKKKs had extensive interaction with WNKN, Mo25, and Mob family proteins, and were involved in plant polarity growth, cell division, and ABA signal transduction. The results of this study might lay a significant foundation for further studies on the gene function and signalling transduction mechanism of MAPKKKK gene family in J. curcas.
引文
Ao PX,Li ZG,Gong M(2013).Involvement of compatible solutes in chill hardening-induced chilling tolerance in Jatropha curcas seedlings.Acta Physiol Plant,35:3457-3464
Bogre L,Meskiene I,Heberle-Bors E,et al(2000).Stressing the role of the MAP kinases in mitogenic stimulation.Plant Mol Biol,43:705-718
?ak?r B,K?l??kaya O(2015).Mitogen-activated protein kinase cascades in Vitis vinifera.Front Plant Sci,6:556
Camps M,Nichols A,Arkinstall S(1999).Dual specificity phosphatases:a gene family for control of MAP kinase function.FASEB J,14:6-16
Champion A,Picaud A,Henry Y(2004).Reassessing the MAP3K and MAP4K relationships.Trends Plant Sci,9:123-129
Chuang HC,Wang XH,Tan TH(2015).MAP4K family kinases in immunity and inflammation.Adv immunol,129:277-314
Dan I,Watanabe NM,Kusumi A(2001).The Ste20 group kinases as regulators of MAP kinase cascades.Trends Cell Biol,11:220-230
Delpire E(2009).The mammalian family of sterile 20p-like protein kinases.Pflug Arch Eur J Phy,458:953-967
Frideman A,Perrimon N(2006).High-throughput approaches to dissecting MAPK signaling pathways.Methods,40:262-271
Gu LK,Liu YK,Zong ZJ,et al(2010).Overexpression of maize mitogen-activated protein kinase gene,ZmSIMK1in Arabidopsis increases tolerance to salt stress.Mol Biol Rep,37:4067-4073
Hamel LP,Nicole MC,Sritubtim S,et al(2006).Ancient signals:comparative genomics of plant MAPK and MAPKKgene families.Trends Plant Sci,11:192-198
Hanks SK,Quinn AM,Hunter T(1988).The protein kinase family:conserved features and deduced phylogeny of the catalytic domains.Science,241:42-52
Ichimura K,Shinozaki K,Tena G(2002).Mitogen-activated protein kinase cascades in plants:a new nomenclature.Trends Plant Sci,7:301-308
Lin J,Zhou XW,Tang KX,et al(2004).A survey of the studies on the resources of Jatropha curcas L..J Trop Subtrop Bot,12:285-290(in Chinese with English abstract)[林娟,周选围,唐克轩等(2004).麻疯树植物资源研究概况.热带亚热带植物学报,12:285-290]
Major G,Daigle C,Stafford-Richard T,et al(2009).Characterization of ScMAP4K1,a MAP kinase kinase kinase kinase in volved in ovule,seed and fruit developmentin Solanum chacoense Bitt.Richard,7:181-183
Makkar HPS,Becker K(2009).Jatropha curcas,a promising crop for the generation of biodiesel and value-added coproducts.Eur J Lipid Sci Tech,111:773-787
Pedley KF,Martin GB(2005).Role of mitogen-activated protein kinase in plant immunity.Curr Opin Plant Biol,8:541-547
Popescu SC,Popescu GV,Bachan S,et al(2009).MAPKtarget networks in Arabidopsis thaliana revealed using functional protein microarrays.Gene Dev,23:80-92
Sato S,Hirakawa H,Isobe S,et al(2011).Sequence analysis of the genome of an oil-bearing tree,Jatropha curcas L..DNA Res,18:65-76
Strange K,Denton J,Nehrke K(2006).Ste20-type kinases:Evolutionarily conserved regulators of ion transport and cell volume.Physiology,21:61-68
Wang HB,Gong M,Guo JY,et al(2018).Genome-wide identification of Jatropha curcas MAPK,MAPKK,and MAP-KKK gene families and their expression profile under cold stress.Sci Rep,8:16163
Wang L,Liu Y,Cai GH,et al(2014).Ectopic expression of ZmSIMK1 leads to improved drought tolerance and activation of systematic acquired resistance in transgenic tobacco.J Biotechnol,172:18-29
Wu PZ,Zhou CP,Cheng SF,et al(2015).Integrated genome sequence and linkage map of physic nut(Jatropha curcas L.),a biodiesel plant.Plant J,81:810-821
Yang CY,Fang Z,Li B,et al(2012).Review and prospects of Jatropha biodiesel industry in China.Renew Sust Energ Rev,16:2178-2190
Zhang L,He LL,Fu QT,et al(2013).Selection of reliable reference genes for gene expression studies in the biofuel plant Jatropha curcas using real-time quantitative PCR.Int J Mol Sci,14:24338-24354
Bogre L,Meskiene I,Heberle-Bors E,et al(2000).Stressing the role of the MAP kinases in mitogenic stimulation.Plant Mol Biol,43:705-718
?ak?r B,K?l??kaya O(2015).Mitogen-activated protein kinase cascades in Vitis vinifera.Front Plant Sci,6:556
Camps M,Nichols A,Arkinstall S(1999).Dual specificity phosphatases:a gene family for control of MAP kinase function.FASEB J,14:6-16
Champion A,Picaud A,Henry Y(2004).Reassessing the MAP3K and MAP4K relationships.Trends Plant Sci,9:123-129
Chuang HC,Wang XH,Tan TH(2015).MAP4K family kinases in immunity and inflammation.Adv immunol,129:277-314
Dan I,Watanabe NM,Kusumi A(2001).The Ste20 group kinases as regulators of MAP kinase cascades.Trends Cell Biol,11:220-230
Delpire E(2009).The mammalian family of sterile 20p-like protein kinases.Pflug Arch Eur J Phy,458:953-967
Frideman A,Perrimon N(2006).High-throughput approaches to dissecting MAPK signaling pathways.Methods,40:262-271
Gu LK,Liu YK,Zong ZJ,et al(2010).Overexpression of maize mitogen-activated protein kinase gene,ZmSIMK1in Arabidopsis increases tolerance to salt stress.Mol Biol Rep,37:4067-4073
Hamel LP,Nicole MC,Sritubtim S,et al(2006).Ancient signals:comparative genomics of plant MAPK and MAPKKgene families.Trends Plant Sci,11:192-198
Hanks SK,Quinn AM,Hunter T(1988).The protein kinase family:conserved features and deduced phylogeny of the catalytic domains.Science,241:42-52
Ichimura K,Shinozaki K,Tena G(2002).Mitogen-activated protein kinase cascades in plants:a new nomenclature.Trends Plant Sci,7:301-308
Lin J,Zhou XW,Tang KX,et al(2004).A survey of the studies on the resources of Jatropha curcas L..J Trop Subtrop Bot,12:285-290(in Chinese with English abstract)[林娟,周选围,唐克轩等(2004).麻疯树植物资源研究概况.热带亚热带植物学报,12:285-290]
Major G,Daigle C,Stafford-Richard T,et al(2009).Characterization of ScMAP4K1,a MAP kinase kinase kinase kinase in volved in ovule,seed and fruit developmentin Solanum chacoense Bitt.Richard,7:181-183
Makkar HPS,Becker K(2009).Jatropha curcas,a promising crop for the generation of biodiesel and value-added coproducts.Eur J Lipid Sci Tech,111:773-787
Pedley KF,Martin GB(2005).Role of mitogen-activated protein kinase in plant immunity.Curr Opin Plant Biol,8:541-547
Popescu SC,Popescu GV,Bachan S,et al(2009).MAPKtarget networks in Arabidopsis thaliana revealed using functional protein microarrays.Gene Dev,23:80-92
Sato S,Hirakawa H,Isobe S,et al(2011).Sequence analysis of the genome of an oil-bearing tree,Jatropha curcas L..DNA Res,18:65-76
Strange K,Denton J,Nehrke K(2006).Ste20-type kinases:Evolutionarily conserved regulators of ion transport and cell volume.Physiology,21:61-68
Wang HB,Gong M,Guo JY,et al(2018).Genome-wide identification of Jatropha curcas MAPK,MAPKK,and MAP-KKK gene families and their expression profile under cold stress.Sci Rep,8:16163
Wang L,Liu Y,Cai GH,et al(2014).Ectopic expression of ZmSIMK1 leads to improved drought tolerance and activation of systematic acquired resistance in transgenic tobacco.J Biotechnol,172:18-29
Wu PZ,Zhou CP,Cheng SF,et al(2015).Integrated genome sequence and linkage map of physic nut(Jatropha curcas L.),a biodiesel plant.Plant J,81:810-821
Yang CY,Fang Z,Li B,et al(2012).Review and prospects of Jatropha biodiesel industry in China.Renew Sust Energ Rev,16:2178-2190
Zhang L,He LL,Fu QT,et al(2013).Selection of reliable reference genes for gene expression studies in the biofuel plant Jatropha curcas using real-time quantitative PCR.Int J Mol Sci,14:24338-24354
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