Franklin A J, Jetton T L, Shelton K D, Magnuson M A
Department of Molecular Physiology and Biophysics, Vanderbilt University Medical School, Nashville, Tennessee 37232.
Mol Cell Biol. 1994 Oct;14(10):6773-88. doi: 10.1128/mcb.14.10.6773-6788.1994.
We report the fortuitous isolation of cDNA clones encoding a novel zinc finger DNA-binding protein termed BZP. The protein encoded is 114 kDa and contains eight zinc finger motifs, seven of which are present in two clusters at opposite ends of the molecule. Both finger clusters bound to the 9-bp sequence AAAGGTGCA with apparent Kds of approximately 2.5 nM. Two of the finger motifs within the amino- and carboxy-terminal finger clusters share 63% amino acid identity. BZP inhibited transcription of the herpes simplex virus thymidine kinase promoter when copies of the 9-bp target motif were linked in cis, suggesting that it functions as a transcriptional repressor. BZP mRNA and immunoreactivity were detected in several established cell lines but were most abundant in hamster insulinoma (HIT) cells, the parental source of the cDNAs. In mouse tissues, BZP mRNA and immunoreactivity were identified in cells of the endocrine pancreas, anterior pituitary, and central nervous system. Interestingly, in HIT cells proliferating in culture, BZP immunoreactivity was predominately nuclear in location, whereas it was usually located in the cytoplasm in most neural and neuroendocrine tissues. Serum deprivation of HIT cells caused BZP immunoreactivity to become predominantly cytoplasmic in location and attenuated its inhibitory effect on transcription, thereby suggesting that the both the subcellular location and the function of this protein are modulated by factors in serum.
我们报告了偶然分离出的编码一种名为BZP的新型锌指DNA结合蛋白的cDNA克隆。所编码的蛋白质为114 kDa,含有八个锌指基序,其中七个位于分子两端的两个簇中。两个指簇均与9碱基序列AAAGGTGCA结合,表观解离常数约为2.5 nM。氨基末端和羧基末端指簇中的两个指基序具有63%的氨基酸同一性。当9碱基靶基序的拷贝顺式连接时,BZP抑制单纯疱疹病毒胸苷激酶启动子的转录,表明它作为转录抑制因子发挥作用。在几种已建立的细胞系中检测到BZP mRNA和免疫反应性,但在仓鼠胰岛素瘤(HIT)细胞(cDNA的亲本来源)中最为丰富。在小鼠组织中,在内分泌胰腺、垂体前叶和中枢神经系统的细胞中鉴定出BZP mRNA和免疫反应性。有趣的是,在培养中增殖的HIT细胞中,BZP免疫反应性主要位于细胞核中,而在大多数神经和神经内分泌组织中它通常位于细胞质中。血清剥夺HIT细胞导致BZP免疫反应性主要位于细胞质中,并减弱其对转录的抑制作用,从而表明该蛋白的亚细胞定位和功能均受血清中因子的调节。