辣椒小G蛋白CaRab11基因全长cDNA的分离及序列分析
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摘要
通过分析克隆获得的辣椒Rab11全长cDNA及其编码的氨基酸序列结构特征,为进一步研究辣椒Rab11功能奠定基础。通过对辣椒均一化cDNA文库的筛选分离获得了一个与葡萄Rab11小G蛋白VvRabA1f高度同源的全长cDNA,命名为CaRab11。序列分析结果表明:该cDNA包含有1 164 bp的完整开放阅读框,编码217个氨基酸。该蛋白含有Rab GTP酶超家族保守的RabSF模体;Rab亚家族蛋白所特有的五个氨基酸序列RabF模体(RabF1-RabF5);参与GTP/Mg++结合及GTP水解的G结构域(G1-G5:GDSGVGKS,T,DTAGQE,GNKADL及ETSAL);两个构象变构域(SwitchⅠ和SwitchⅡ)及与异戊二烯化相关的CCX序列。氨基酸同源性及进化分析同样表明CaRab11为辣椒小G蛋白Rab11家族新成员。
The characteristics of Rab11 cDNA and deduced amino acids sequence in pepper were analyzed,which would lay the foundation for further investigation of the function of Rab11 protein.One 1 164 bp full-length cDNA clone was isolated from a pepper normalized cDNA library,which encodes a putative protein composed of 217 amino acids.The full-length cDNA was named CaRab11.The amino acid sequence deduced by CaRab11 cDNA showed high similarity to VvRabA1f protein from Vitis vinifera.CaRab11 protein included conserved domains specific to Rab GTPase superfamily(RAbsF),Rab subfamily specific regions(RabF,RabF1-RabF5),five highly conserved GTP-binding consensus sequences(GDSGVGKS,T,DTAGQE,GNKADL,and ETSAL) involved in GTP/ Mg++ binding and GDP hydrolysis,two conformational switch(Switch I and II) regions as well as C-terminal cysteines motif important to prenylation.Amino acids similarity and phylogenetic analysis also indicated that CaRab11 is a new member of small GTP-binding protein Rab11 superfamily.
The characteristics of Rab11 cDNA and deduced amino acids sequence in pepper were analyzed,which would lay the foundation for further investigation of the function of Rab11 protein.One 1 164 bp full-length cDNA clone was isolated from a pepper normalized cDNA library,which encodes a putative protein composed of 217 amino acids.The full-length cDNA was named CaRab11.The amino acid sequence deduced by CaRab11 cDNA showed high similarity to VvRabA1f protein from Vitis vinifera.CaRab11 protein included conserved domains specific to Rab GTPase superfamily(RAbsF),Rab subfamily specific regions(RabF,RabF1-RabF5),five highly conserved GTP-binding consensus sequences(GDSGVGKS,T,DTAGQE,GNKADL,and ETSAL) involved in GTP/ Mg++ binding and GDP hydrolysis,two conformational switch(Switch I and II) regions as well as C-terminal cysteines motif important to prenylation.Amino acids similarity and phylogenetic analysis also indicated that CaRab11 is a new member of small GTP-binding protein Rab11 superfamily.
引文
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[16]赖燕,肖翔,林箐,等.辣椒小G蛋白CaRab8基因全长cDNA的分离及表达特征的初步分析[J].热带作物学报,2011,32(6):1111-1115.
[17]Yim Y S,Moak P,Sanchez-Villeda H,et al.A BAC pooling strategy combined with PCR-based screenings in a large,highly repetitive genome enables integration of the maize genetic and physical maps[J].BMC Genomics,2007,8:47.
[18]Vernoud V,Horton A C,Yang Z,et al.Analysis of the small GTPase gene superfamily of Arabidopsis[J].Plant Physiol2003,131(3):1191-1208.
[2]Novick P,Zerial M.The diversity of Rab proteins in vesicle transport[J].Curr Opin Cell Biol,1997,9(4):496-504.
[3]Pereira-Leal J B,Seabra M C.The mammalian Rab family of small GTPases:definition of family and subfamily sequencemotifs suggests a mechanism for functional specificity in the Ras superfamily[J].J Mol Biol,2000,301(4):1077-1087.
[4]Molendijk A J,Ruperti B,Palme K.Small GTPases in vesicle trafficking[J].Curr Opin Plant Biol,2004,7(6):694-700.
[5]Surpin M,Raikhel N.Traffic jams affect plant development and signal transduction[J].Nat Rev Mol Cell Biol,2004,5(2):100-109.
[6]Bischoff F,Molendijk A,Rajendrakumar C S,et al.GTP-binding proteins in plants[J].Cell Mol Life Sci,1999,55(2):233-256.
[7]Stenmark H,Olkkonen V M.The Rab GTPase family[J].Genome Biol,2001,2(5):3007.
[8]Volpicelli L A,Lah J J,Fang G,et al.Rab11a and myosin Vb regulate recycling of the M4 muscarinic acetylcholine recep-tor[J].J Neurosci,2002,22(22):9776-9784.
[9]Schlierf B,Fey G H,Hauber J,et al.Rab11b is essential for recycling of transferrin to the plasma membrane[J].Exp CellRes,2000,259(1):257-265.
[10]Wilcke M,Johannes L,Galli T,et al.Rab11 regulates the compartmentalization of early endosomes required for efficienttransport from early endosomes to the trans-golgi network[J].J Cell Biol,2000,151(6):1207-1220.
[11]Lu C,Zainal Z,Tucker G A,et al.Developmental abnormalities and reduced fruit softening in tomato plants expressing anantisense Rab11 GTPase gene[J].Plant Cell,2001,13(8):1819-1833.
[12]De Graaf B H,Cheung A Y,Andreyeva T,et al.Rab11 GTPase-regulated membrane trafficking is crucial for tip-fo-cused pollen tube growth in tobacco[J].Plant Cell,2005,17(9):2564-2579.
[13]Nagano Y,Okada Y,Narita H,et al.Location of light-repressible,small GTP-binding protein of the YPT/rab family inthe growing zone of etiolated pea stems[J].Proc Natl Acad Sci USA,1995,92(14):6314-6318.
[14]Kang J G,Yun J,Kim D H,et al.Light and brassinosteroid signals are integrated via a dark-induced small G protein inetiolated seedling growth[J].Cell,2001,105(5):625-636.
[15]徐小万,李颖,王恒明.中国辣椒工业的现状、发展趋势及对策[J].园艺园林科学,2008,24(11):332-338.
[16]赖燕,肖翔,林箐,等.辣椒小G蛋白CaRab8基因全长cDNA的分离及表达特征的初步分析[J].热带作物学报,2011,32(6):1111-1115.
[17]Yim Y S,Moak P,Sanchez-Villeda H,et al.A BAC pooling strategy combined with PCR-based screenings in a large,highly repetitive genome enables integration of the maize genetic and physical maps[J].BMC Genomics,2007,8:47.
[18]Vernoud V,Horton A C,Yang Z,et al.Analysis of the small GTPase gene superfamily of Arabidopsis[J].Plant Physiol2003,131(3):1191-1208.