|
|
|
|
|
| Screening and Analysis of Proteins Interacting with TaPDK from Physiological Male Sterility Induced by CHA in Wheat |
| ZHANG Long-yu, ZHANG Gai-sheng, ZHAO Xin-liang , YANG Shu-ling |
| College of Agronomy, Northwest A&F University/Wheat Breeding Engineering Research Center, Ministry of Education/Key Laboratory of Crop Heterosis of Shaanxi Province, Yangling 712100, P.R.China |
|
|
|
摘要 To further research the regulatory network of pyruvate dehydrogenase kinase (designated as TaPDK) in physiological male-sterility (PHYMS) of wheat induced by chemical hybridizing agent (CHA) SQ-1, an anther cDNA library was constructed, and the proteins interacting with TaPDK were screened via yeast two-hybrid technique. Subsequently, a few candidate proteins in nucleotide expression levels were detected by real-time quantitative PCR. Yeast-two hybrid screening was performed by mating yeast strain Y2HGold containing BD-TaPDK bait plasmid with yeast strain Y187 including anther cDNA library plasmid. Diploid yeast cells were plated on synthetic dropout nutrient medium (SD/-Ade/ -His/-Leu/-Trp) (QDO), and further were incubated on QDO medium containing AbA and X-α-Gal. The interactions between TaPDK and the proteins obtained from positive colonies were further confirmed by co-transformation validation. After plasmids DNA were extracted from blue colonies and sequenced, the sequences results were analyzed by bioinformatic methods. Finally, 24 colonies were obtained, including eight genes, namely non-specific lipid-transfer protein precursor (TanLTP), polyubiquitin (TaPUbi), glyceraldehyde-3-phosphate dehydrogenase, proliferating cell nuclear antigen (TaPCNA), CBS domain containing protein (TaCBS), actin, guanine nucleotide-binding protein beta subunit, chalcone synthase, and three new genes with unknown function. The results of quantitative RT-PCR showed that the expression levels of TanLTP, TaPUbi, and TaPCNA were obviously up-regulated in PHYMS anther, and TaCBS expression was only increased at the tricellular stage in PHYMS anther compared with in fertile lines. Whereas, the expression of TaPDK was obviously down-regulated in PHYMS lines. Collectively, these datas indicated that the majority of candidate proteins might be related to pollen abortion in PHYMS lines, which further suggested that TaPDK plays multiple roles in pollen development, besides participating in regulating pyruvate dehydrogenase complex activity.
Abstract To further research the regulatory network of pyruvate dehydrogenase kinase (designated as TaPDK) in physiological male-sterility (PHYMS) of wheat induced by chemical hybridizing agent (CHA) SQ-1, an anther cDNA library was constructed, and the proteins interacting with TaPDK were screened via yeast two-hybrid technique. Subsequently, a few candidate proteins in nucleotide expression levels were detected by real-time quantitative PCR. Yeast-two hybrid screening was performed by mating yeast strain Y2HGold containing BD-TaPDK bait plasmid with yeast strain Y187 including anther cDNA library plasmid. Diploid yeast cells were plated on synthetic dropout nutrient medium (SD/-Ade/ -His/-Leu/-Trp) (QDO), and further were incubated on QDO medium containing AbA and X-α-Gal. The interactions between TaPDK and the proteins obtained from positive colonies were further confirmed by co-transformation validation. After plasmids DNA were extracted from blue colonies and sequenced, the sequences results were analyzed by bioinformatic methods. Finally, 24 colonies were obtained, including eight genes, namely non-specific lipid-transfer protein precursor (TanLTP), polyubiquitin (TaPUbi), glyceraldehyde-3-phosphate dehydrogenase, proliferating cell nuclear antigen (TaPCNA), CBS domain containing protein (TaCBS), actin, guanine nucleotide-binding protein beta subunit, chalcone synthase, and three new genes with unknown function. The results of quantitative RT-PCR showed that the expression levels of TanLTP, TaPUbi, and TaPCNA were obviously up-regulated in PHYMS anther, and TaCBS expression was only increased at the tricellular stage in PHYMS anther compared with in fertile lines. Whereas, the expression of TaPDK was obviously down-regulated in PHYMS lines. Collectively, these datas indicated that the majority of candidate proteins might be related to pollen abortion in PHYMS lines, which further suggested that TaPDK plays multiple roles in pollen development, besides participating in regulating pyruvate dehydrogenase complex activity.
|
|
Received: 17 September 2012
Accepted:
|
| Fund: Acknowledgements The research was supported by the National High-Tech R&D Program of China (2011AA10A106), the National Natural Science Foundation of China (31071477, 31171611), and the Key Scientific and Technological Innovation Special Projects of Shaanxi “13115”, China (2010ZDKG-68, 2011KTZB02-01-01). |
|
Corresponding Authors:
Correspondence ZHANG Gai-sheng, Tel: +86-29-87092152, Fax: +86-29-87092292, E-mail: zhanggsh@public.xa.sn.cn
E-mail: zhanggsh@public.xa.sn.cn
|
Cite this article:
ZHANG Long-yu, ZHANG Gai-sheng, ZHAO Xin-liang , YANG Shu-ling.
2013.
Screening and Analysis of Proteins Interacting with TaPDK from Physiological Male Sterility Induced by CHA in Wheat. Journal of Integrative Agriculture, 12(6): 941-950.
|
| [1]Ariizumi T, Amagai M, Shibata D, Hatakeyama K,Watanabe M, Toriyama K. 2002. Comparative study ofpromoter activity of three anther-specific genesencoding lipid transfer protein, xyloglucane n d o t r a n s g l u c o s y l a s e / h y d r o l a s e a n dpolygalacturonase in transgenic Arabidopsis thaliana.Plant Cell Report, 21, 90-96[2]Bateman A. 1997. The structure of a domain common toarchaebacteria and the homocystinuria disease protein.Trends in Biochemical Sciences, 22, 12-13[3]Baxi M D, Vishwanatha J K. 1995. Uracil DNA-glycosylase/glyceraldehyde-3-phosphate dehydrogenase is anAp4A binding protein[4]Biochemistry, 34, 9700-9707[5]Binet N N, Wei J H, Tessier L H. 1991. Structure andexpression of sunflower ubiquitin genes. PlantMolecular Biology, 17, 395-470[6]Boutrot F, Meynard D, Guiderdoni E, Joudrier P, Gautier MF. 2007. The Triticum aestivum non-specific lipid transferprotein (TaLtp) gene family: comparative promoteractivity of six TaLtp genes in transgenic rice. Planta,225, 843-862[7]Bravo R, Fey S J, Bellatin J, Larsen P M, Celis J E. 1982.Identification of nuclear polypeptide (‘cyclin’) whoserelative proportion is sensitive to changes in the rate ofcell proliferation and to transformation. Progress inClinical and Biological Research, 85, 235-248[8]Bryksin A V, Laktionov P P. 2008. Role of glyceraldehyde-3-phosphate dehydrogenase in vesicular transport fromgolgi apparatus to endoplasmic reticulum[9]Biochemistry,73, 619-625[10]Budde R J A, Randall D D. 1990. Pea leaf mitochondrialpyruvate dehydrogenase complex is inactivated in vivoin a light-dependent manner. Proceedings of theNational Academy of Sciences of the United States ofAmerica, 8, 673-676[11]Butterfield D A, Hardas S S, Lange M L. 2010. Oxidativelymodified glyceraldehyde-3-phosphate dehydrogenase(GAPDH) and alzheimer disease: many pathways toneurodegeneration Journal of Alzheimers Disease, 20,369-393[12]Christensen A H, Sharrok RA, Quail P H. 1992. Maizepolyubiquitin genes: structure, thermal perturbation ofexpression and transcript splicing, and promoter activityfollowing transfer to protoplasts by electroporation.Plant Molecular Biology, 18, 675-689[13]Durrieu C, Bernier-Valentin F, Rousset B. 1987. Binding ofglyceraldehyde-3-phosphate dehydrogenase tomicrotubules Molecular Cell Biochemistry, 74, 55-65[14]Engel M, Seifert M, Theisinger B, Seyfert U, Welter C. 1998.Glyceraldehyde-3-phosphate dehydrogenase andNm23-H1/nucleoside diphosphate kinase A Journalof Biological Chemistry, 273, 20058-20065[15]González-Melendi P, Testillano P S, Ahmadian P, Reyes J,Risueño M C. 2000. Immunoelectron microscopy of PCNA as an efficient marker for studying replicationtimes and sites during pollen development.Chromosoma, 109, 397-409[16]Haglund K, Dikic I. 2005. Ubiquitylation and cell signaling.EMBO Journal, 24, 3353-3359[17]Higaki T, Kojo K H, Hasezawa S. 2010. Critical role of actinbundling in plant cell morphogenesis. Plant Signaling& Behavior, 5, 484-488[18]Hoffman N E, Ko K, Milkowski D, Pichersky E. 1991.Isolation and characterization of tomato cDNA andgenomic clones encoding the ubiquitin gene ubi3. PlantMolecular Biology, 17, 1189-1201[19]Huang B, Wu P M, Bowker-Kinley M M, Robert A, Harris RA. 2002. Regulation of pyruvate dehydrogenase kinaseexpression by peroxisome proliferator-activatedreceptor-??ligands, glucocorticoids, and insulin.Diabetes, 51, 276-283[20]Ignoul S, Eggermont J. 2005. CBS domains: structure,function, and pathology in human proteins. AmericanJournal of Physiology (Cell Physiology), 289, C1369-C1378.Kader J C. 1996. Lipid-transfer proteins in plants.Annual Review of Plant Physiology and Plant MolecularBiology, 47, 627-654[21]Kawalleck P, Somssich I E, Feldbrugge M, Hahlbrock K,Weisshaar B. 1993. Polyubiquitin gene expression andstructural properties of the ubi4-2 gene in Petroseliumcrispum Plant Molecular Biology, 21, 673-684[22]Kim T H, Park J H, Kim M C, Cho S H. 2008. Cutin monomerinduces expression of the rice OsLTP5 lipid transferprotein gene. Journal of Plant Physiology, 165, 345-349[23]Lee S B, Go Y S, Bae H J, Park J H, Cho S H, Cho H J, Lee DS, Park O K, Hwang I, Suh M C. 2009. Disruption ofglycosylphosphatidylinositol-anchored lipid transferprotein gene altered cuticular lipid composition,increased plastoglobules, and enhanced susceptibilityto infection by the fungal pathogen alternariabrassicicola. Plant Physiology, 150, 42-54[24]Lernmark U, Gardestrom P. 1994. Distribution of pyruvatedehydrogenase complex activities between chloroplastsand mitochondria from leaves of different species. PlantPhysiology, 106, 1633-1638[25]Li R J, Hu Z Y, Zhang H S, Zhan G M, Wang H Z, Hua W.2011. Cloning and functions analysis of a pyruvatedehydrogenase kinase in Brassica napus. Plant CellReports, 30, 1533-1540[26]Liu H W, Zhang G S, Wang J W, Wang X L, Fang Z W.2003. Effect of male sterility on different wheat genotypeinduced by SQ-1 Journal of Northwest A & F University(Natural Science Edition), 31, 15-18[27](in Chinese)Livak K J, Schmittgen T D. 2001. Analysis of relative geneexpression data using real-time quantitative PCR andthe 2-??Ct method. Methods, 25, 402-408[28]Ma L G, Xu X D, Cui S J, Sun D Y. 1999. The presence of aheterotrimeric G protein and its role in signaltransduction of extracellular calmodulin in pollendevelopment germination and tube growth. The PlantCell, 11, 1351-1363[29]Meyer-Siegler K, Mauro D J, Seal G, Wurzer J, deRiel J K,Sirover M A. 1991. A human nuclear uracil DNAglycosylase is the 37-kDa subunit of glyceraldehyde-3-phosphate dehydrogenase Proceedings of theNational Academy of Sciences of the United States ofAmerica, 88, 8460-8464[30]Naryzhny S N. 2008. Proliferating cell nuclear antigen: aproteomics view. Cellular and Molecular LifeSciences, 65, 3789-3808[31]Park S Y, Jauh G Y, Mollet J C, Eckard K J, Nothnagel E A,Walling L L, Lord E M. 2000. A lipid transfer-like proteinis necessary for lily pollen tube adhesion to an in vitrostylar matrix. Plant Cell Physiology, 12, 151-164[32]Raynaud C, Sozzani R, Glab N, Domenichini S, Perennes C,Cella R, Kondorosi E, Bergounioux C. 2006. Two cellcycleregulated SET-domain proteins interact withproliferating cell nuclear antigen (PCNA) inArabidopsis. The Plant Journal, 47, 395-407[33]Peškan-Berghöfer T, Neuwirth J, Kusnetsov V, OelmüllerR. 2005. Suppression of heterotrimeric G-protein betasubunitaffects anther shape, pollen development andinflorescence architecture in tobacco. Planta, 220, 737-746[34]Sarowar S, Kim Y J, Kim K D, Hwang B K, Ok S H, Shin J S.2009. Over-expression of lipid transfer protein (LTP)genes enhances resistance to plant pathogens and LTPfunctions in long-distance systemic signaling intobacco. Plant Cell Report, 28, 419-427[35]Schijlen E G W M, Vos C H R D, Martens S, Jonker H H,Rosin F M, Molthoff J W, Tikunov Y M, Angenent G C,Tunen A J V, Bovy A G. 2007. RNA interference silencingof chalcone synthase, the first step in the flavonoidbiosynthesis pathway, leads to parthenocarpic tamatofruits. Plant Physiology, 144, 1520-1530[36]Scorah J, Dong M Q, Yates III J R, Scott M, Gillespie D,McGowan C H. 2008. A conserved proliferating cellnuclear antigen-interacting protein sequence in Chk1is required for checkpoint function. Journal ofBiological Chemistry, 283, 17250-17259[37]Simon M I, Strathmann M P, Gautam N. 1991. Diversity of Gproteins in signal transduction. Science, 252, 802-808[38]Smalle J, Vierstra R D. 2004. The ubiquitin 26S proteasomeproteolytic pathway. Annual Review of Plant Biology,55, 555-590[39]Sun C L, Pan Y Y. 2008. Testing pollen character duringpollen development in Arabidopsis. Chinese Bulletinof Botany, 25, 268-275[40](in Chinese)Tedege M, Kuhlemeier C. 1997. Aerobic fermentation duringtobacco pollen development. Plant Molecular Biology,35, 3433-3454[41]Thompson P, Reid E E, Lyttle C R, Dennis D T. 1977.Pyruvate dehydrogenase complex from higher plantmitochondria and proplastids: regulation. PlantPhysiology, 59, 854-858[42]Tover-Mendez A T, Miernyk J A, Randall D D. 2003.Regulation of pyruvate dehydrogenase complex activityin plant cells. European Journal of Biochemistry, 270,1043-1049[43]Volker K W, Knull H. 1997. A glycolytic enzyme bindingdomain on tubulin. Archives of BiochemistryBiophysics, 338, 237-243[44]Wei M M,W S J, Zhang G H, Zhang L Y, Yuan Z J, Sun R, YeJ X, Niu N, Ma S C, Li H X. 2009. Relationship betweenthe expression of GAPDH gene and anther abortion ofphysiological male sterile of wheat. Molecular PlantBreeding, 7, 679-684[45](in Chinese)Williams M, Randall D D. 1979. Pyruvate dehydrogenasecomplex from chloroplasts of Pisum sativum L. PlantPhysiology, 64, 1099-1103[46]Xu C H, Liu Z T, Zhang L P, Zhao C P, Yuan S H, Zhang FT. 2013. Organization of actin cytoskeleton duringmeiosis I in a wheat thermo-sensitive genic male sterileline. Protoplasma, 250, 415-422[47]Yamada M. 1992, Lipid transfer proteins in plants andmicroorganisms. Plant Cell Physiology, 33, 1-6[48]Yan L F, Liu G Q, Xiao X G. 2000. From pollen actin to cropmale sterility. Chinese Science Bulletin, 45, 784-788[49]Yang M, Liu X H, Huang J B. 2008. Study on flavonoidsextraction of microwave method and ethanol from lotusleaves. Storage and Process, 8, 31-33[50]Yui R, Iketani S, Mikami T, Kubo T. 2003. Atisense inhibitionof mitochondrial pyruvate dehydrogenase E1??subunitin anther tapetum causes male sterility. The PlantJournal, 34, 57-66[51]Zhang D S, Liang W Q, Yin C, Zong J, Gu F W, Zhang D B.2010a. OsC6, encoding a lipid transfer protein, isrequired for postmeiotic anther development in rice.Plant Physiology, 154, 149-162[52]Zhang L Y, Li H X, Zhang G, Wang J, Han Y F, Yuan Z J, NiuN, Ma S C. 2009. Cloning and expression analysis ofcMDH gene related to cytoplasmic male sterile wheatwith Aegilops kotschyi Cytoplasm. Acta AgronomicaSinica, 35, 1620-1627[53](in Chinese)Zhang L Y, Yuan L, Yang S L, Zhang G S. Wang J S, Song YL, Zhao Z J, Niu N, Ma S C. 2011. Expressioncharacteristic of TaPDC-E1??gene and its regulatoryenzymes gene in male sterile line of wheat. ActaAgronomica Sinica, 37, 620-628 (in Chinese)[54]Zhang M Z, Yuan L, Wang J S, Zhang G S, Guo W J, ZhangL Y. 2010. Expression analysis of cACO gene in malesterile wheat induced by chemical hybridzing agentSQ-1 Chinese Journal of Biochemistry and MolecularBiology, 26, 740-748 (in Chinese)[55]Zheng L, Roeder R G, Luo Y. 2003. S-phase activation ofthe histone H2B promoter by OCA-S, a coactivatorcomplex that contains GAPDH as a key component.Cell, 114, 255-266[56]Zou J T, Qi Q G, Katavic V, Marillia E F, Taylor D C. 1999.Effects of antisense repression of an Arabidopsisthaliana pyruvate dehydrogenase kinase cDNA onplant development. Plant Molecular Biology, 41, 837-849 |
| No Suggested Reading articles found! |
|
|
Viewed |
|
|
|
Full text
|
|
|
|
|
Abstract
|
|
|
|
|
Cited |
|
|
|
|
| |
Shared |
|
|
|
|
| |
Discussed |
|
|
|
|
