Choi Hee-Jung, Chung Tae-Wook, Kang Nam-Young, Kim Kyoung-Sook, Lee Young-Choon, Kim Cheorl-Ho
Faculty of Biotechnology, Dong-A University, Busan 604-714, South Korea.
FEBS Lett. 2003 Dec 4;555(2):204-8. doi: 10.1016/s0014-5793(03)01227-4.
We studied the transcriptional regulation of human GM3 synthase (hST3Gal V) during monocytic differentiation of HL-60 cells induced by 12-O-tetradecanoylphorbol-13-acetate (TPA). Northern blot and reverse transcription polymerase chain reaction indicated that the induction of hST3Gal V by TPA is regulated at the transcriptional level. To elucidate the mechanism underlying the regulation of hST3Gal V gene expression during the differentiation of HL-60 cells induced by TPA, we characterized the promoter region of the hST3Gal V gene. Functional analysis of the 5'-flanking region of the hST3Gal V gene by the transient expression method showed that the -177 to -83 region, which contains a cAMP responsive element binding protein (CREB) binding site at -143, functions as the TPA-inducible promoter in HL-60 cells. In addition, gel shift assays and site-directed mutagenesis indicated that the CREB binding site at -143 is crucial for the TPA-induced expression of the hST3Gal V in HL-60 cells.
我们研究了在12 - O - 十四酰佛波醇 - 13 - 乙酸酯（TPA）诱导的HL - 60细胞单核细胞分化过程中人类GM3合酶（hST3Gal V）的转录调控。Northern印迹和逆转录聚合酶链反应表明，TPA对hST3Gal V的诱导是在转录水平上调控的。为了阐明TPA诱导的HL - 60细胞分化过程中hST3Gal V基因表达调控的潜在机制，我们对hST3Gal V基因的启动子区域进行了特征分析。通过瞬时表达方法对hST3Gal V基因5' - 侧翼区域进行功能分析表明，-177至-83区域（在-143处含有一个环磷酸腺苷反应元件结合蛋白（CREB）结合位点）在HL - 60细胞中作为TPA诱导型启动子发挥作用。此外，凝胶迁移试验和定点诱变表明，-143处的CREB结合位点对于TPA诱导的HL - 60细胞中hST3Gal V的表达至关重要。
