Gutierrez-Hartmann A, Lieberburg I, Gardner D, Baxter J D, Cathala G G
Nucleic Acids Res. 1984 Sep 25;12(18):7153-73. doi: 10.1093/nar/12.18.7153.
The rat growth hormone (rGH) gene contains two classes of repetitive DNA arranged as clusters within intron B and the 3' flanking region. The major family is equivalent to the CHO type 2 DNA. The second ("truncated repeat", TR) is a truncated version of the first and occurs in certain neural-specific transcripts and genes ("identifier" elements, ID). Here we report, using the HeLa cell-free transcription assay, that RNA polymerase III (Pol III) efficiently initiates at internal promoters within a tandem array of rGH gene repetitive DNA monomers and results in a novel organization of overlapping Class III transcription units. Transcription competition studies revealed that the rat type 2 structures share Pol III transcription factors with a tRNA gene, a human Alu repeat, and a mutant VA1 gene. Also, the rGH type 2 but not the TR DNA efficiently promotes Pol III initiation, yet other TR members, which differ only in flanking DNA, are transcribed. Thus, the rGH gene is strikingly enriched with 10 repetitive DNA monomers; multimeric type 2 elements are actively transcribed; rGH-TR sequences are expressed only as part of larger transcripts promoted by type 2 DNA; and, type 2 DNA uses tRNA gene transcription factors. These studies show that flanking sequences, promoter organization and factor competition may all affect rat repetitive DNA expression.
大鼠生长激素(rGH)基因包含两类重复DNA,它们以内含子B和3'侧翼区域内的簇状形式排列。主要家族等同于CHO 2型DNA。第二种(“截短重复序列”,TR)是第一种的截短版本,存在于某些神经特异性转录本和基因(“标识符”元件,ID)中。在此,我们使用HeLa无细胞转录试验报告,RNA聚合酶III(Pol III)能在rGH基因重复DNA单体串联阵列内的内部启动子处高效起始转录,并导致重叠的III类转录单元形成一种新的组织形式。转录竞争研究表明,大鼠2型结构与tRNA基因、人类Alu重复序列和突变的VA1基因共享Pol III转录因子。此外,rGH 2型而非TR DNA能有效促进Pol III起始转录,然而,仅侧翼DNA不同的其他TR成员也会被转录。因此,rGH基因显著富集了10个重复DNA单体;多聚体2型元件能被积极转录;rGH - TR序列仅作为由2型DNA促进的较大转录本的一部分而表达;并且,2型DNA使用tRNA基因转录因子。这些研究表明,侧翼序列、启动子组织和因子竞争可能都会影响大鼠重复DNA的表达。