ISOLATION AND CHARACTERIZATION OF A NOVEL CDNA FROM HL-60 CELLS TREATED WITH 1,25-DIHYDROXYVITAMIN D-3
ISOLATION AND CHARACTERIZATION OF A NOVEL CDNA FROM HL-60 CELLS TREATED WITH 1,25-DIHYDROXYVITAMIN D-3
复制标题
DOI：
10.1016/0167-4781(94)90242-9
复制
发表时间：
1994-09-13
期刊：
影响因子：
--
通讯作者：
DELUCA, HF
中科院分区：
文献类型：
--
作者：
CHEN, KS;DELUCA, HF
从1，25-二羟基维生素D-3刺激的HL-60细胞的mRNA文库中分离到一个新的cDNA克隆(VDUP1)。Northern印迹分析表明，VDUP1基因与2.9kb的mRNA杂交，在1，25-二羟基维生素D-3(1，25-(OH)(2)D-3)作用下，HL-60细胞中VDUP1基因表达上调。VDUP1基因的体外表达产生了一个46 kDa的蛋白。对GenBank数据库的搜索显示，VDUP1的3‘非翻译区与在脑中表达的序列同源。详细的时程研究表明，1，25-二羟基维生素D-3作用6h后，VDUP1mRNA开始升高，18h左右达到高峰，并持续升高至24h，且不受佛波酯(PMA)调节。环己亚胺抑制蛋白质合成并不能阻止1，25-二羟基维生素D-3对VDUP1mRNA的诱导。放线菌酮本身可增加VDUP1的mRNA水平。这些结果表明，VDUP1 mRNA的降解要么是翻译依赖的，要么是由不稳定的蛋白质调节的。
A novel cDNA clone (VDUP1) has been isolated from a cDNA library constructed from mRNA obtained from HL-60 cells stimulated by 1,25-dihydroxyvitamin D-3. Northern blot analysis showed that VDUP1 cDNA hybridizes to a 2.9 kb mRNA species which is up-regulated in HL-60 cells by 1,25-dihydroxyvitamin D-3 (1,25-(OH)(2)D-3) treatment. In vitro expression of VDUP1 cDNA produced a 46 kDa protein. A search of the GenBank database revealed that the 3' untranslated region of VDUP1 is homologous to a sequence expressed in brain. A detailed time course study showed that the VDUP1 mRNA starts to increase at 6 h after 1,25-dihydroxyvitamin D-3 treatment, reaches a plateau at around 18 h and stays elevated for 24 h. The VDUP1 mRNA is not regulated by phorbol 12-myristate 13-acetate (PMA) in HL-60 cells. Inhibition of protein synthesis by cycloheximide does not prevent the induction of VDUP1 mRNA by 1,25-dihydroxyvitamin D-3. Cycloheximide itself increases VDUP1 mRNA levels. These results suggest that the degradation of VDUP1 mRNA is either translation-dependent or regulated by a labile protein.