Kunz D, Zimmermann R, Heisig M, Heinrich P C
Biochemisches Institut, Universität Freiburg, FRG.
Nucleic Acids Res. 1989 Feb 11;17(3):1121-38. doi: 10.1093/nar/17.3.1121.
The 5'-region of the rat alpha 2-macroglobulin gene has been characterized. A 5.6 kb Sal I - Xba I fragment containing the first 4 exons of the alpha 2-macroglobulin gene and 1.3 kb of its 5'-flanking region was sequenced. The putative transcriptional start site was determined by RNase protection and primer extension analysis. TATA- and CAAT-box equivalent sequences were found. A potential glucocorticoid receptor binding site was located on the antisense strand. DNA sequences containing the 5'-flanking region of the rat alpha 2-macroglobulin gene were linked to the gene coding for the bacterial chloramphenicol acetyltransferase and introduced into Hep G2 cells. In these transfected Hep G2 cells CAT activity could be induced by recombinant human interleukin-6. Deletion analyses have shown that the sequences between -852 and -777 as well as between -404 and -165 relative to the cap site, contain regulatory elements involved in the interleukin-6 dependent induction of the alpha 2-macroglobulin gene.
大鼠α2-巨球蛋白基因的5'区域已得到鉴定。对一个5.6 kb的Sal I - Xba I片段进行了测序，该片段包含α2-巨球蛋白基因的前4个外显子及其5'侧翼区域的1.3 kb。通过核糖核酸酶保护和引物延伸分析确定了推定的转录起始位点。发现了类似TATA盒和CAAT盒的序列。在反义链上定位了一个潜在的糖皮质激素受体结合位点。将含有大鼠α2-巨球蛋白基因5'侧翼区域的DNA序列与编码细菌氯霉素乙酰转移酶的基因相连，并导入Hep G2细胞。在这些转染的Hep G2细胞中，重组人白细胞介素-6可诱导氯霉素乙酰转移酶（CAT）活性。缺失分析表明，相对于帽位点，-852至-777以及-404至-165之间的序列包含参与白细胞介素-6依赖性诱导α2-巨球蛋白基因的调控元件。
