Welcome to Acta Agronomica Sinica,

Acta Agron Sin ›› 2014, Vol. 40 ›› Issue (12): 2081-2089.doi: 10.3724/SP.J.1006.2014.02081

• CROP GENETICS & BREEDING·GERMPLASM RESOURCES·MOLECULAR GENETICS • Previous Articles     Next Articles

Screening of Promoter-Binding Factors of Tobacco PMT Gene Using a Modified Yeast Surface Display System

CHEN Hong,NIU Hai-Xia,WANG Wen-Jing,MA Hao-Ran,LI Jia-Na,CHAI You-Rong,ZHANG Hong-Bo   

  1. College of Agronomy and Biotechnology, Southwest University/Chongqing Engineering Research Center for Rapeseed, Chongqing 400716, China
  • Received:2014-05-15 Revised:2014-09-16 Online:2014-12-12 Published:2014-10-08
  • Contact: 张洪博, E-mail: hbzhang@swu.edu.cn

Abstract:

Yeast surface display system is an important tool for studying molecular interaction of proteins, however, its application in DNA-binding-protein screening is relatively limited. Yeast surface display system secretes exogenous proteins onto cell surface, thus, it could be applied to determine protein interaction under in-vitro-like experiment conditions. Therefore, it may be more efficient than yeast one-hybrid system in screening the DNA-binding proteins whose DNA-binding capability is affected by endogenous yeast factors. In this study, we modified the pYD1 vector in yeast surface display system to make it compatible with the Smart cDNA library construction kit from Clontech, which will be helpful to increase the library construction efficiency. An experimental procedure for DNA-binding protein isolation was established using the modified yeast surface display system. Then, it was successfully applied in screening DNA-binding proteins of a jasmonate (JA) responsive element in the promoter of tobacco PMT (putrescine N-methyltransferase) gene. Among the isolated genes, two encoded ERF transcription factors, which were found to bind the JA responsive element in PMT promoter in vitro but unable to activate the expression of reporters in yeast-one-hybrid system. Our study suggests that yeast surface display system is efficient in screening the DNA-binding proteins whose DNA-binding capability in yeast-one-hybrid system is disrupted by endogenous factors.

Key words: Yeast surface display, Smart cDNA Library, DNA-binding protein, Yeast one-hybrid, PMT gene

[1]Schwechheimer C, Zourelidou M, Bevan M W. Plant transcription factor studies. Annu Rev Plant Physiol Plant Mol Biol, 1998, 49: 127–150
[2]廖名湘, 方福德. 酵母单杂交体系--一种研究DNA-蛋白质相互作用的有效方法. 中国医学科学院学报, 2000, 22: 388–391
Liao M X, Fang F D. Yeast one-hybrid system--one effective method studying DNA-protein interaction. Acta Acad Med Sin, 2000, 22: 388–391 (in Chinese with English abstract)
[3]王佳堃, 孙中远, 刘建新. 酵母细胞表面展示技术. 动物营养学报, 2011, 23: 1847–1853
Wang J K, Sun Z Y, Liu J X. Recent advances in yeast cell-surface display technology. Chin J Anim Nutr, 2011, 23: 1847–1853 (in Chinese with English abstract)
[4]Kondo A, Ueda M. Yeast cell-surface display--applications of molecular display. Appl Microbiol Biotechnol, 2004, 64: 28–40
[5]Pepper L R, Cho Y K, Boder E T, Shusta E V. A decade of yeast surface display technology: where are we now? Comb Chem High Throughput Screen, 2008, 11: 127–134
[6]Gera N, Hussain M, Rao B M. Protein selection using yeast surface display. Methods, 2013, 60: 15–26
[7]Boder E T, Raeeszadeh-Sarmazdeh M, Price J V. Engineering antibodies by yeast display. Arch Biochem Biophys, 2012, 526: 99–106
[8]罗立新, 吴琳, 林影. 酵母表面展示分选酶底物用于分选酶活性检测. 微生物学报, 2009, 49: 1534–1539
Luo L X, Wu L, Lin Y. Studies on the sortase activity assay via display its substrates on yeast surface. Acta Microb Sin, 2009, 49: 1534–1539 (in Chinese with English abstract)
[9]张伟, 郭钦, 阮辉, 张洪波, 何国庆. 酵母表面展示技术在蛋白质工程中的应用. 生物技术通报, 2009, (8): 63–66
Zhang W, Guo Q, Ruan H, Zhang H B, He G Q. Application of yeast cell-surface display for protein engine application of yeast cell-surface display for protein engineering. Biotechnol Bull, 2009, (8): 63–66 (in Chinese with English abstract)
[10]郭钦, 张伟, 阮晖, 何国庆. 酿酒酵母表面展示表达系统及应用. 中国生物工程杂志, 2008, 28 (12): 116–122
Guo Q, Zhang W, Ruan H, He G Q. Cell-surface display expression system of Saccharomyces cerevisiae and its applications. Chin Biotechnol, 2008, 28 (12): 116–122 (in Chinese with English abstract)
[11]Lofblom J. Bacterial display in combinatorial protein engineering. Biotechnol J, 2011, 6: 1115–1129
[12]Bratkovic T. Progress in phage display: evolution of the technique and its application. Cell Mol Life Sci, 2010, 67: 749–767
[13]Tanaka T, Yamada R, Ogino C, Kondo A. Tanaka T, Yamada R, Ogino C, Kondo A. Recent developments in yeast cell surface display toward extended applications in biotechnology. Appl Microbiol Biotechnol, 2012, 95: 577–591
[14]Bidlingmaier S, Wang Y, Liu Y, Zhang N, Liu B. Comprehensive analysis of yeast surface displayed cDNA library selection outputs by exon microarray to identify novel protein-ligand interactions. Mol Cell Proteomics, 2011, 10: M110 005116
[15]Chattopadhyay M K, Ghosh B. Molecular analysis of polyamine biosynthesis in higher plants. Curr Sci, 1998, 74: 517–522
[16]Chou W M, Kutchan T M. Enzymatic oxidations in the biosynthesis of complex alkaloids. Plant J, 1998, 15: 289–300
[17]Riechers D E, Timko M P. Structure and expression of the gene family encoding putrescine N-methyltransferase in Nicotiana tabacum: new clues to the evolutionary origin of cultivated tobacco. Plant Mol Biol, 1999, 41: 387–401
[18]Shoji T, Yamada Y, Hashimoto T. Jasmonate induction of putrescine N-methyltransferase genes in the root of Nicotiana sylvestris. Plant Cell Physiol, 2000, 41: 831–839
[19]Baldwin I T. Jasmonate-induced responses are costly but benefit plants under attack in native populations. Proc Natl Acad Sci USA, 1998, 95: 8113–8118
[20]Imanishi S, Hashizume K, Nakakita M, Kojima H, Matsubayashi Y, Hashimoto T, Sakagami Y, Yamada Y, Nakamura K. Differential induction by methyl jasmonate of genes encoding ornithine decarboxylase and other enzymes involved in nicotine biosynthesis in tobacco cell cultures. Plant Mol Biol, 1998, 38: 1101–1111
[21]Shoji T, Hashimoto T. Why does anatabine, but not nicotine, accumulate in jasmonate-elicited cultured tobacco BY-2 cells? Plant Cell Physiol, 2008, 49: 1209–1216
[22]Xu B, Timko M. Methyl jasmonate induced expression of the tobacco putrescine N-methyltransferase genes requires both G-box and GCC-motif elements. Plant Mol Biol, 2004, 55: 743–761
[23]Sears M T, Zhang H, Rushton P J, Wu M, Han S, Spano A J, Timko M P. NtERF32: a non-NIC2 locus AP2/ERF transcription factor required in jasmonate-inducible nicotine biosynthesis in tobacco. Plant Mol Biol, 2013, 84:49–66
[24]Zhang H B, Bokowiec M T, Rushton P J, Han S C, Timko M P. Tobacco transcription factors NtMYC2a and NtMYC2b form nuclear complexes with the NtJAZ1 repressor and regulate multiple jasmonate-inducible steps in nicotine biosynthesis. Mol Plant, 2012, 5: 73–84
[25]De Sutter V, Vanderhaeghen R, Tilleman S, Lammertyn F, Vanhoutte I, Karimi M, Inzé D, Goossens A, Hilson P. Exploration of jasmonate signaling via automated and standardized transient expression assays in tobacco cells. Plant J, 2005, 44: 1065–1076
[26]Todd A T, Liu E, Polvi S L, Pammett R T, Page J E. A functional genomics screen identifies diverse transcription factors that regulate alkaloid biosynthesis in Nicotiana benthamiana. Plant J, 2010, 62: 589–600
[27]Shoji T, Ogawa T, Hashimoto T. Jasmonate-induced nicotine formation in tobacco is mediated by tobacco COI1 and JAZ genes. Plant Cell Physiol, 2008, 49: 1003–1012
[1] WANG Zhen, ZHANG Xiao-Li, MENG Xiao-Jing, YAO Meng-Nan, MIU Wen-Jie, YUAN Da-Shuang, ZHU Dong-Ming, QU Cun-Min, LU Kun, LI Jia-Na, LIANG Ying. Identification of upstream regulators for mitogen-activated protein kinase 7 gene (BnMAPK7) in rapeseed (Brassica napus L.) [J]. Acta Agronomica Sinica, 2021, 47(12): 2379-2393.
[2] JIA Shuang-Wei,GAO Ying,ZHAO Kai-Jun. Cloning and Characterization of Brassica juncea Zinc Finger Protein Transcription Factor Gene Bj26 [J]. Acta Agron Sin, 2014, 40(07): 1174-1181.
Viewed
Full text


Abstract

Cited

  Shared   
  Discussed   
No Suggested Reading articles found!