Tajima Hiromi, Iwata Yuji, Iwano Megumi, Takayama Seiji, Koizumi Nozomu
Graduate school of Biological Sciences, Nara Institute of Science and Technology, 8916-5 Takayama, Ikoma, Nara 630-0192, Japan.
Biochem Biophys Res Commun. 2008 Sep 19;374(2):242-7. doi: 10.1016/j.bbrc.2008.07.021. Epub 2008 Jul 15.
Among 75 bZIP transcription factors identified in Arabidopsis, 3 (AtbZIP17, AtbZIP28, and AtbZIP49) possess a putative transmembrane domain (TMD) in addition to AtbZIP60, which was characterized previously. In the present study, cDNAs of AtbZIP17 and AtbZIP28 were isolated. Truncated forms of AtbZIP17 and AtbZIP28 lacking the C-terminal domain including TMD were examined as putative active forms. One of them, AtbZIP28DeltaC, activated BiP1 and BiP3 promoters through the cis-elements P-UPRE and ERSE responsible for the ER stress response. Subsequently, a fusion protein of green fluorescent protein (GFP) and AtbZIP28 was expressed in Arabidopsis cultured cells. Under non-stress conditions, GFP fluorescence localization almost overlapped with an ER marker; however, tunicamycin and dithiothreitol treatment clearly increased GFP fluorescence in the nucleus suggesting that the N-terminal fragment of AtbZIP28 translocates to the nucleus in response to ER stress.
在拟南芥中鉴定出的75个bZIP转录因子中,除了先前已被表征的AtbZIP60外,还有3个(AtbZIP17、AtbZIP28和AtbZIP49)具有一个推定的跨膜结构域(TMD)。在本研究中,分离出了AtbZIP17和AtbZIP28的cDNA。对缺失包括TMD在内的C末端结构域的AtbZIP17和AtbZIP28的截短形式作为推定的活性形式进行了检测。其中之一,AtbZIP28DeltaC,通过负责内质网应激反应的顺式元件P-UPRE和ERSE激活了BiP1和BiP3启动子。随后,绿色荧光蛋白(GFP)与AtbZIP28的融合蛋白在拟南芥培养细胞中表达。在非应激条件下,GFP荧光定位几乎与内质网标记物重叠;然而,衣霉素和二硫苏糖醇处理明显增加了细胞核中的GFP荧光,表明AtbZIP28的N末端片段在响应内质网应激时易位至细胞核。