Ito Naoko, Takabatake Reona, Seo Shigemi, Hiraga Susumu, Mitsuhara Ichiro, Ohashi Yuko
Core Research of Evolutionary Science and Technology, 2-3 Surugadai, Kannda, Chiyoda-Ku, Tokyo, 101-0062, Japan.
Plant Cell Physiol. 2002 Mar;43(3):266-74. doi: 10.1093/pcp/pcf031.
A gene encoding a receptor-like protein kinase was isolated as the gene induced in the early period of N gene-dependent hypersensitive cell death in tobacco leaves. The kinase domain expressed as a glutathione S-transferase fusion protein was capable of autophosphorylation, indicating that this gene encodes an active protein kinase. A high level of the transcript accumulated before necrotic lesion formation in tobacco mosaic virus (TMV)-inoculated tobacco leaves carrying the N gene but it was low in a tobacco cultivar lacking the N gene. A small but reproducible increase in the transcript was found 1-2 h after a temperature shift from 30 degrees C to 20 degrees C even in healthy leaves, suggesting the gene expression is temperature sensitive. The gene was named WRK for wound-induced receptor-like protein kinase, because the transcript increased to a maximum within 15-30 min of wounding. In suspension cultured tobacco cells, an increase in the transcript was found 15 min after transfer to a new medium, but it was suppressed under high osmotic pressures. The wound-induced WRK accumulation was enhanced by cycloheximide treatment, but not by known defense signal compounds (salicylic acid, jasmonic acid, 1-aminocyclopropan-1-carboxylic acid and abscisic acid) and some plant hormones. Thus, WRK is a wound-inducible and temperature-sensitive protein kinase gene induced before hypersensitive cell death probably through unknown signaling pathways.
一个编码类受体蛋白激酶的基因被分离出来,它是在烟草叶片中N基因依赖的超敏细胞死亡早期被诱导的基因。以谷胱甘肽S-转移酶融合蛋白形式表达的激酶结构域能够进行自身磷酸化,这表明该基因编码一种活性蛋白激酶。在携带N基因的烟草花叶病毒(TMV)接种的烟草叶片中,在坏死病斑形成之前积累了高水平的转录本,但在缺乏N基因的烟草品种中该转录本水平较低。即使在健康叶片中,从30℃温度转移到20℃后1-2小时也发现转录本有小但可重复的增加,这表明基因表达对温度敏感。该基因被命名为WRK,即伤口诱导的类受体蛋白激酶,因为转录本在受伤后15-30分钟内增加到最大值。在悬浮培养的烟草细胞中,转移到新培养基后15分钟发现转录本增加,但在高渗透压下受到抑制。伤口诱导的WRK积累通过环己酰亚胺处理增强,但不受已知防御信号化合物(水杨酸、茉莉酸、1-氨基环丙烷-1-羧酸和脱落酸)和一些植物激素的影响。因此,WRK是一个伤口诱导且对温度敏感的蛋白激酶基因,可能通过未知信号通路在超敏细胞死亡之前被诱导。