Grichnik J M, French B A, Schwartz R J
Department of Cell Biology, Baylor College of Medicine, Houston, Texas 77030.
Mol Cell Biol. 1988 Nov;8(11):4587-97. doi: 10.1128/mcb.8.11.4587-4597.1988.
The chicken skeletal alpha-actin gene promoter region (-202 to -12) provides myogenic transcriptional specificity. This promoter contains partial dyad symmetry about an axis at nucleotide -108 and in transfection experiments is capable of directing transcription in a bidirectional manner. At least three different transcription initiation start sites, oriented toward upstream sequences, were mapped 25 to 30 base pairs from TATA-like regions. The opposing transcriptional activity was potentiated upon the deletion of sequences proximal to the alpha-actin transcription start site. Thus, sequences which serve to position RNA polymerase for alpha-actin transcription may allow, in their absence, the selection of alternative and reverse-oriented start sites. Nuclear runoff transcription assays of embryonic muscle indicated that divergent transcription may occur in vivo but with rapid turnover of nuclear transcripts. Divergent transcriptional activity enabled us to define the 3' regulatory boundary of the skeletal alpha-actin promoter which retains a high level of myogenic transcriptional activity. The 3' regulatory border was detected when serial 3' deletions bisected the element (-91 CCAAA TATGG -82) which reduced transcriptional activity by 80%. Previously we showed that disruption of its upstream counterpart (-127 CCAAAGAAGG -136) resulted in about a 90% decrease in activity. These element pairs, which we describe as CCAAT box-associated repeats, are conserved in all sequenced vertebrate sarcomeric actin genes and may act in a cooperative manner to facilitate transcription in myogenic cells.
鸡骨骼肌α-肌动蛋白基因启动子区域(-202至-12)具有成肌转录特异性。该启动子在核苷酸-108处围绕一个轴具有部分二重对称,并且在转染实验中能够以双向方式指导转录。至少三个不同的转录起始位点,朝向上游序列,定位在距TATA样区域25至30个碱基对处。在删除α-肌动蛋白转录起始位点近端的序列后,相反的转录活性增强。因此,用于将RNA聚合酶定位用于α-肌动蛋白转录的序列在不存在时可能允许选择替代的和反向定向的起始位点。胚胎肌肉的核径流转录分析表明,体内可能发生分歧转录,但核转录本周转迅速。分歧转录活性使我们能够定义骨骼肌α-肌动蛋白启动子的3'调控边界,该边界保留了高水平的成肌转录活性。当连续的3'缺失将使转录活性降低80%的元件(-91 CCAAA TATGG -82)二等分时,检测到3'调控边界。先前我们表明,其上游对应物(-127 CCAAAGAAGG -136)的破坏导致活性下降约90%。我们将这些元件对描述为CCAAT盒相关重复序列,在所有已测序的脊椎动物肌节肌动蛋白基因中都是保守的,并且可能以协同方式作用以促进成肌细胞中的转录。