Hara K, Yagi M, Kusano T, Sano H
Research and Education Center for Genetic Information, Nara Institute of Science and Technology, Ikoma, Japan.
Mol Gen Genet. 2000 Feb;263(1):30-7. doi: 10.1007/pl00008673.
An immediate-early, transiently activated wound-responsive gene was identified in tobacco by fluorescent differential display screening. The full-length cDNA encodes a polypeptide of 356 amino acids with a relative molecular mass of 39,082 Da. The deduced amino acid sequence shows two characteristic features; a leucine-zipper motif found in the more N-terminal region and a WRKY domain containing a zinc-finger motif located in the central region. The gene was designated as wizz (wound-induced leucine zipper zinc finger). Northern analysis showed that upon wounding wizz transcripts were locally and systemically accumulated within 10 min, reached a maximum level by 30 min, and decreased thereafter to the basal level. Analyses of a WIZZ-GFP fusion protein clearly indicated that WIZZ is a nuclear factor. WIZZ specifically binds to sequences containing two TTGAC core motifs that are separated by a spacer of appropriate length. The binding activity was dependent on bivalent cations, most probably zinc. In transient reporter assays, however, WIZZ did not show transactivation activity in tobacco suspension cells, suggesting that it functions together with other components. The results indicate that WIZZ is a new transcription factor which participates in early stages of the wound response.
通过荧光差异显示筛选在烟草中鉴定出一个即时早期、瞬时激活的伤口响应基因。该全长cDNA编码一个由356个氨基酸组成的多肽,相对分子质量为39,082 Da。推导的氨基酸序列显示出两个特征;在更靠近N端区域发现一个亮氨酸拉链基序,在中央区域有一个包含锌指基序的WRKY结构域。该基因被命名为wizz(伤口诱导的亮氨酸拉链锌指)。Northern分析表明,受伤后wizz转录本在10分钟内局部和系统地积累,30分钟时达到最高水平,此后下降至基础水平。对WIZZ-GFP融合蛋白的分析清楚地表明WIZZ是一种核因子。WIZZ特异性结合含有两个由适当长度间隔区隔开的TTGAC核心基序的序列。结合活性依赖于二价阳离子,很可能是锌。然而,在瞬时报告基因测定中,WIZZ在烟草悬浮细胞中未显示反式激活活性,表明它与其他成分一起发挥作用。结果表明WIZZ是一种参与伤口响应早期阶段的新转录因子。