Spitkovsky D D, Royer-Pokora B, Delius H, Kisseljov F, Jenkins N A, Gilbert D J, Copeland N G, Royer H D
Angewandte Tumorvirologie, Deutsches Krebsforschungszentrum, Heidelberg, FRG.
Nucleic Acids Res. 1992 Feb 25;20(4):797-803. doi: 10.1093/nar/20.4.797.
YB-1 cDNA clones were isolated by binding site screening of a Hela expression library using a human papillomavirus type 18 enhancer oligonucleotide. YB-1 belongs to a family of transcription factors which bind to recognition sequences containing a core CCAAT element. YB-1 bound to its single stranded recognition sequence on the sense strand but not to the anti-sense strand. A synthetic peptide antiserum derived from the predicted YB-1 amino acid sequence identified a 42 kD nuclear protein in immunoblots. A protein with the same size was detected by binding site blotting experiments using the HPV18 enhancer oligonucleotide which bound YB-1. YB-1 gene expression was restricted in tissues from a human 24 week old fetus. High levels of YB-1 mRNA were present in heart, muscle, liver, lung, adrenal gland and the brain, in contrast, low amounts of YB-1 mRNA were found in thymus, kidney, bone marrow and spleen. In pancreas, bladder, stomach and testis YB-1 mRNA could not be detected by Northern hybridization. Finally, we have identified four YB-1 related loci in the mouse genome and have mapped these loci to four different mouse chromosomes by interspecific backcross analysis.
通过使用人乳头瘤病毒18型增强子寡核苷酸对Hela表达文库进行结合位点筛选,分离出YB-1 cDNA克隆。YB-1属于一类转录因子家族,其可与含有核心CCAAT元件的识别序列结合。YB-1可与其正义链上的单链识别序列结合,但不与反义链结合。源自预测的YB-1氨基酸序列的合成肽抗血清在免疫印迹中鉴定出一种42 kD的核蛋白。使用结合YB-1的HPV18增强子寡核苷酸通过结合位点印迹实验检测到一种大小相同的蛋白质。YB-1基因表达在一名24周龄人类胎儿的组织中受到限制。心脏、肌肉、肝脏、肺、肾上腺和大脑中存在高水平的YB-1 mRNA,相反,在胸腺、肾脏、骨髓和脾脏中发现低水平的YB-1 mRNA。在胰腺、膀胱、胃和睾丸中,通过Northern杂交无法检测到YB-1 mRNA。最后,我们在小鼠基因组中鉴定出四个与YB-1相关的基因座,并通过种间回交分析将这些基因座定位到四条不同的小鼠染色体上。