Regulation of rat brain natriuretic peptide transcription. A potential role for GATA-related transcription factors in myocardial cell gene expression.
Regulation of rat brain natriuretic peptide transcription. A potential role for GATA-related transcription factors in myocardial cell gene expression.
复制标题
DOI：
--
复制
发表时间：
1994-07
期刊：
影响因子：
--
通讯作者：
D. Thuerauf;D. S. Hanford;C. Glembotski
中科院分区：
文献类型：
--
作者：
D. Thuerauf;D. S. Hanford;C. Glembotski
GATA结合蛋白是调节红系细胞中珠蛋白基因的阶段特异性和组织特异性表达的转录因子。最近，心脏GATA结合蛋白被发现是心脏发生过程中最早表达的基因，然而，心脏中该转录因子的靶基因尚不清楚。由于脑利钠肽（BNP）在心脏生长发育的早期被激活，我们评估了它是否可以作为GATA结合蛋白介导的诱导的靶基因。在分离和测序大鼠BNP 5 '-侧翼序列（FS）的2.5个内切酶后，鉴定了多种推定的转录增强子位点，包括几个加塔共有序列（WGATAR），其中一个显然充当主要启动子位点。用BNP/荧光素酶融合基因转染原代心肌细胞;典型的生长因子如佛波酯、血清或α 1-肾上腺素能激动剂以及加塔-4过表达强烈诱导报告基因表达。截短分析显示，诱导蛋白主要定位于大鼠BNP 5 '-FS的近端-116碱基对，其中除了TATA盒处的加塔序列外，还有两个共有加塔位点。点突变分析表明，至少有一个加塔位点是赋予完整的加塔-4诱导转录所必需的。这些结果表明，大鼠BNP 5 '-FS的近端区域是生长因子和GATA诱导的转录所必需的，支持BNP基因可以在早期心脏发育期间作为GATA结合蛋白的靶点的观点。
GATA-binding proteins are transcription factors that regulate the stage- and tissue-specific expression of globin genes in cells of the erythroid lineage. Recently, a cardiac GATA-binding protein was found to be the earliest gene expressed during cardiogenesis; however, the target genes of this transcription factor in the heart are unknown. Since brain natriuretic peptide (BNP) is activated early in cardiac growth and development, we evaluated whether it could serve as a target gene for GATA-binding protein-mediated induction. Upon isolating and sequencing 2.5 kilobases of the rat BNP 5'-flanking sequence (FS), a variety of putative transcriptional enhancer sites were identified, including several GATA consensus sequences (WGATAR), one of which apparently serves as the major promoter site. Primary myocardial cells were transfected with BNP/luciferase fusion genes; reporter expression was strongly induced by typical growth factors such as phorbol esters, serum, or alpha 1-adrenergic agonists, as well as by GATA-4 overexpression. Truncation analyses showed that inducibility mapped primarily to the proximal -116 base pair of the rat BNP 5'-FS, where there are two consensus GATA sites in addition to the GATA sequence at the TATA box. Point mutation analyses showed that at least one of the GATA sites was required to confer full GATA-4-inducible transcription. These results demonstrate that a proximal region of the rat BNP 5'-FS is required for growth factor- and GATA-inducible transcription, supporting the view that the BNP gene could serve as a target for GATA-binding proteins during early cardiac development.