Heard D J, Filipowicz W, Marques J P, Palme K, Gualberto J M
Friedrich Miescher-Institut, Basel, Switzerland.
Nucleic Acids Res. 1995 Jun 11;23(11):1970-6. doi: 10.1093/nar/23.11.1970.
The genes transcribed by RNA polymerase (pol) III can be placed into four distinct groups based on the nature and position of their promoter elements. In the higher eukaryotes equivalent genes usually belong to the same sub-type of pol III promoters and there are few examples of genes which have changed promoter type during evolution. In this work we demonstrate that the promoter of the Arabidopsis thaliana 7SL RNA gene is located upstream of the coding region and is identical to the promoters of pol III-specific plant U-small nuclear RNA (U-snRNA) genes. Sequence analysis of two different 7SL genes from A. thaliana revealed that both genes contain two sequence elements in their 5' flanking regions identical in sequence and position to the highly conserved USE and TATA elements of the pol III-transcribed plant U-snRNA genes. Mutational analysis of these elements in the At7SL-2 gene indicates that the USE and TATA elements are both necessary and account for > or = 90% of the transcriptional activity of this gene in transfected plant protoplasts. Within the coding region of both genes there is a sequence element which is a 10/11 nt match to the consensus B-box element of tRNA genes, however, this element is not important for gene activity. These findings distinguish the plant genes from the human 7SL gene, which has both internal and upstream promoter elements and its upstream elements are different from those found in the human U-snRNA genes.
RNA聚合酶(pol)III转录的基因可根据其启动子元件的性质和位置分为四个不同的组。在高等真核生物中,等效基因通常属于pol III启动子的同一亚型,在进化过程中改变启动子类型的基因实例很少。在这项工作中,我们证明拟南芥7SL RNA基因的启动子位于编码区上游,与pol III特异性植物U小核RNA(U-snRNA)基因的启动子相同。对来自拟南芥的两个不同7SL基因的序列分析表明,这两个基因在其5'侧翼区域都含有两个序列元件,其序列和位置与pol III转录的植物U-snRNA基因的高度保守的USE和TATA元件相同。对At7SL-2基因中这些元件的突变分析表明,USE和TATA元件都是必需的,并且在转染的植物原生质体中占该基因转录活性的≥90%。在这两个基因的编码区内有一个序列元件,它与tRNA基因的共有B-box元件有10/11个核苷酸匹配,然而,这个元件对基因活性并不重要。这些发现将植物基因与人类7SL基因区分开来,人类7SL基因既有内部启动子元件又有上游启动子元件,其上游元件与人类U-snRNA基因中的不同。