Stimac E, Lyons S, Pious D
Department of Pediatrics, University of Washington, Seattle 98195.
Mol Cell Biol. 1988 Sep;8(9):3734-9. doi: 10.1128/mcb.8.9.3734-3739.1988.
HLA-DR and other human class II histocompatibility genes are expressed by Epstein-Barr virus-transformed B-lymphocyte cell lines but not by most T-cell leukemia lines. We determined by transcriptional run-on experiments that regulation of class II expression in these cells is at the level of gene transcription; nuclei isolated from B-cell lines actively transcribe class II mRNA, whereas nuclei from non-class II-expressing T-cell lines and from the class II transactive factor-deficient B-cell mutant 6.1.6 do not. In searching for DNA-binding proteins which might regulate transcription, we found both a ubiquitous (B1) and a B-cell-specific (B2) factor which bind to the octamer sequence ATTTGCAT 52 base pairs 5' of the cap site in the DR alpha gene. We examined the relationship of these factors to DR alpha transcription. HUT-78, a T-cell line which expresses class II mRNA constitutively, contains only the ubiquitous B1 octamer-binding factor also found in non-class II-expressing T-cell leukemias. Human fibroblast, HeLa, and melanoma cell lines similarly contain only the ubiquitous factor, even when these cells are induced to express class II mRNA by treatment with gamma interferon. Both B1 and B2 binding factors are present in the B-cell mutant 6.1.6, which nevertheless fails to transcribe class II mRNA. Although we have not ruled out the requirement of B-cell-specific octamer-binding factor B2 for class II expression in B cells, it is clear that in other cells substantial DR alpha transcription occurs in the absence of this factor.
HLA - DR及其他人类Ⅱ类组织相容性基因可由爱泼斯坦 - 巴尔病毒转化的B淋巴细胞系表达,但大多数T细胞白血病系则不表达。我们通过转录连续实验确定,这些细胞中Ⅱ类基因表达的调控处于基因转录水平;从B细胞系分离的细胞核可积极转录Ⅱ类mRNA,而从不表达Ⅱ类的T细胞系以及Ⅱ类反式激活因子缺陷的B细胞突变体6.1.6分离的细胞核则不能。在寻找可能调控转录的DNA结合蛋白时,我们发现了一种普遍存在的(B1)和一种B细胞特异性的(B2)因子,它们与DRα基因帽位点5'端52个碱基对处的八聚体序列ATTTGCAT结合。我们研究了这些因子与DRα转录的关系。HUT - 78是一种组成性表达Ⅱ类mRNA的T细胞系,只含有在不表达Ⅱ类的T细胞白血病中也能发现的普遍存在的B1八聚体结合因子。人成纤维细胞、HeLa细胞和黑色素瘤细胞系同样只含有普遍存在因子,即使这些细胞经γ干扰素处理后被诱导表达Ⅱ类mRNA也是如此。B1和B2结合因子都存在于B细胞突变体6.1.6中,但该突变体仍无法转录Ⅱ类mRNA。虽然我们尚未排除B细胞特异性八聚体结合因子B2对B细胞中Ⅱ类表达所必需,但很明显在其他细胞中,即使没有该因子也会发生大量的DRα转录。