CHARACTERIZATION OF THE DECAY-ACCELERATING FACTOR GENE PROMOTER REGION
CHARACTERIZATION OF THE DECAY-ACCELERATING FACTOR GENE PROMOTER REGION
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DOI：
10.1073/pnas.88.11.4675
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发表时间：
1991-06-01
影响因子：
11.1
通讯作者：
MEDOF, ME
中科院分区：
文献类型：
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作者：
EWULONU, UK;RAVI, L;MEDOF, ME
衰变加速因子（Decay-accelerating factor，ERF）表达调节细胞对自体补体攻击的易感性。为了表征控制p53基因转录的调控区，克隆并测序了从815个碱基对（bp）上游延伸到p53 AUG密码子（指定为+1）下游几乎等于4个碱基对的基因组DNA。5'侧翼序列显示59-76%的G+C含量（-355至+1），至少一个GC盒（-135至-131），以及符合基序TCCTCC和TC（n）的可变长度序列（从-629至-285）。核酸酶S1双端测序和引物延伸将主要转录起始位点定位于可能的TATA变体（A）TTTAA下游的-82/-81，38 bp。在COS细胞转染中，包含-815至-67的序列的功能效率为劳斯肉瘤病毒3'长末端重复序列的2.5%，但在-355上游缺失后，其活性增加几乎等于4倍。检测到与佛波醇12-肉豆蔻酸酯13-乙酸酯（PMA）和cAMP响应元件（分别为PRE和克雷斯）具有部分同源性的两个八核苷酸，并且各自的调节剂分别将转录效率提高2倍和几乎等于10倍。因此，该基因启动子（i）显示类似于常规和非常规转录控制元件的序列，（ii）具有负调节活性的区域，和（iii）可能通过PRE-和CRE-样增强子元件响应PMA和cAMP诱导。
Decay-accelerating factor (DAF) expression modulates susceptibility of cells to autologous complement attack. To characterize the regulatory region controlling DAF gene transcription, genomic DNA extending from 815 base pairs (bp) upstream to almost-equal-to 4 kilobases downstream of DAF's AUG codon (designated +1) was cloned and sequenced. The 5' flanking sequence showed 59-76% G+C content (-355 to +1), at least one GC box(es) (-135 to -131), and variable length sequences (from -629 to -285) conforming to the motifs TCCTCC and TC(n). Nuclease S1 digestions and primer extensions localized a major transcriptional start site to -82/ -81, 38 bp downstream of a possible TATA variant, (A)TTTAA. In COS cell transfections, the sequence encompassing -815 to -67 functioned 2.5% as efficiently as the Rous sarcoma virus 3' long terminal repeat, but following deletion upstream of -355 its activity increased almost-equal-to 4-fold. Two octanucleotides exhibiting partial homology to phorbol 12-myristate 13-acetate (PMA) and cAMP responsive elements (PREs and CREs, respectively) were detected, and the respective modulators enhanced transcriptional efficiency 2- and almost-equal-to 10-fold, respectively. Thus, the DAF gene promoter (i) exhibits sequences resembling both conventional and unconventional transcriptional control elements, (ii) possesses a region with negative regulatory activity, and (iii) responds to PMA and cAMP induction presumably via PRE- and CRE-like enhancer elements.