锌指蛋白是一类重要的转录因子, 广泛参与植物的生长发育和胁迫应答过程。文章使用生物信息学方法从水稻中预测并克隆了一个TFⅢA型锌指蛋白基因ZFP207(GenBank 登陆号: AK063147.1), 该基因包含一个567 bp的开放阅读框, 编码一条188个氨基酸组成的多肽,其编码产物的预测分子量为20.72 kDa, 等电点为9.67。锌指蛋白ZFP207含有一个典型的TFⅢA型锌指结构, 并且在C端含有一个可能具有转录抑制功能的EAR-motif, 但不具有任何已知的核定位信号。系统发生分析结果表明, ZFP207与植物中已经发现的单锌指蛋白亲缘关系较近。通过RT-PCR方法分析了ZFP207基因在成株期水稻不同组织中的表达, 发现ZFP207在茎和叶中表达量较高, 而在穗和根中表达量较低。在酵母中的转录激活实验结果显示, ZFP207在酵母中不具有转录激活功能。
The zinc finer proteins consist of a large transcription factor family involved in plant development and responses to environmental stresses. In this paper, a TFⅢA-type zinc finger protein gene ZFP207 (GenBank assession number AK063147.1) was cloned from rice variety Jiucaiqing by RT-PCR approach. This gene contains an open reading frame (ORF) of 567 bp, which encodes a peptide of 188 amino acid residues. The isoelectric point (pI) of the protein is 9.67, and its molecular weight is 20.72 kDa. Bioinformatic analysis showed that the ZFP207 protein comprises a typical TFⅢA-type zinc finger domain and an EAR-motif at its C-terminus. However, nuclear localization signals (NLS) commonly existing in TFⅢA-type zinc finger proteins was not found in the ZFP207 amino acid sequence. In addition, based on the alignments of the whole amino acid sequences of some known TFⅢA-type zinc finger proteins in plants, a phylogenetic tree was con-structed by the neighbour joining method. The phylogenetic tree showed that ZFP207 and other TFⅢA-type zinc finger proteins with single zinc finger domain were grouped into the same branch. The expression pattern of ZFP207 gene was also investigated in various rice tissues at adult stage by RT-PCR and the results showed that ZFP207 was expressed with high levels in culms and leaves, but lower in roots and spikes. Finally, the trans-activation assay in yeast cells revealed that ZFP207 lacked the trans-activation activity.
[1] 刘强, 张贵友, 陈受宜. 植物转录因子的结构与调控作用. 科学通报, 2000, 45(14): 1465–1474.
[2] Takatsuji H. Zinc-finger transcription factors in plants. Cell Mol Life Sci, 1998, 54(6): 582–596.
[3] Takatsuji H, Mori M, Benfey PN, Ren L, Chua NH. Characterization of a zinc finger DNA-binding protein expressed specifically in Petunia petals and seedlings. EMBO J,1992, 11(1): 241–249.
[4] 黄骥, 王建飞, 张红生. 植物C2H2型锌指蛋白的结构与功能. 遗传, 2004, 26(3): 414–418.
[5] Huang J, Yang X, Wang MM, Tang HJ, Ding LY, Shen Y, Zhang HS. A novel rice C2H2-type zinc finger protein lacking DLN-box/EAR-motif plays a role in salt tolerance. Biochim Biophys Acta, 2007, 1769(4): 220–227.
[6] 郭书巧, 黄骥, 江燕, 张红生. 水稻C2H2型锌指蛋白基因RZF71的克隆与表达分析. 遗传, 2007, 29(5): 607–613.
[7] Xu DQ, Huang J, Guo SQ, Yang X, Bao YM, Tang HJ, Zhang HS. Overexpression of a TFIIIA-type zinc finger protein gene ZFP252 enhances drought and salt tolerance in rice (Oryza sativa L.). FEBS Lett, 2008, 582(7): 1037–1043.
[8] Huang J, Wang JF, Wang QH, Zhang HS. Identification of a rice zinc finger protein whose expression is transiently induced by drought, cold but not by salinity and abscisic acid. DNA Seq, 2005, 16(2): 130–136.
[9] 江燕. 水稻和拟南芥TFⅢA型锌指蛋白基因家族的鉴定及ZFP207基因的功能研究[学位论文]. 南京农业大学, 2007.
[10] Yoshida S, Forno DA, Cock JH, Gomez KA. Routine procedure for growing rice plants in culture solution. In: Laboratory Manual for Physiological Studies of Rice, IRRI. Los Banos, the Philippines, 1976, 53–57.
[11] Reece KS, McElroy D, Wu R. Genomic nucleotide se-quence of four rice (Oryza sativa L.) actin genes. Plant Mol Biol, 1990, 14 (4): 621–624.
[12] Ciftci-Yilmaz S, Morsy MR, Song L, Coutu A, Krizek BA, Lewis MW, Warren D, Cushman J, Connolly EL,Mittler R. The EAR-motif of the Cys2/His2-type zinc finger protein Zat7 plays a key role in the defense response of arabidopsis to salinity stress. J Biol Chem, 2007, 282(12): 9260–9268.
[13] Sugano S, Kaminaka H, Rybka Z, Catala R, Salinas J, Matsui K, Ohme-Takagi M, Takatsuji H. Stress-responsive zinc finger gene ZPT2-3 plays a role in drought tolerance in petunia. Plant J, 2003, 36(6): 830–841.
[14] Huang F, Chi Y, Meng Q, Gai J, Yu D. GmZFP1 encoding a single zinc finger protein is expressed with enhancement in reproductive organs and late seed development in soybean (Glycine max). Mol Biol Rep, 2006, 33(4): 279–285.
[15] Agarwal P, Arora R, Ray S, Singh AK, Singh VP, Takatsuji H, Kapoor S, Tyagi AK. Genome-wide identification of C2H2 zinc-finger gene family in rice and their phylogeny and expression analysis. Plant Mol Biol, 2007, 65(4): 467–485.
[16] Hiratsu K, Ohta M, Matsui K, Ohme-Takagi M. The SUPERMAN protein is an active repressor whose carboxy-terminal repression domain is required for the development of normal flowers. FEBS Lett, 2002, 514(2): 351–354.
[17] Kim SH, Hong JK, Lee SC, Sohn KH, Jung HW, Hwang BK. CAZFP1, Cys2/His2-type zinc-finger transcription factor gene functions as a pathogen-induced early-defense gene in Capsicum annuum. Plant Mol Biol, 2004, 55(6): 883–904.