Dahlberg J E, Schenborn E T
Department of Physiological Chemistry, University of Wisconsin-Madison 53706.
Nucleic Acids Res. 1988 Jul 11;16(13):5827-40. doi: 10.1093/nar/16.13.5827.
We examined the ability of the 5' flanking region sequences of a human U1 RNA gene to direct synthesis of functional mRNA. When fused to chloramphenicol acetyltransferase (CAT) coding region sequences, the upstream sequences of the U1 gene were able to stimulate the synthesis of functional CAT mRNA in 293 cells but not in HeLa cells. Most of the polyadenylated CAT mRNA in 293 cells originated from cryptic promoters in the upstream U1 sequences, but nearly all of the CAT-specific RNA originating at position +1 (relative to the U1 gene promoter) was non-polyadenylated; this confirmed that the bona-fide U1 gene promoter was unable to direct efficient synthesis of poly-A+ mRNA. Our results demonstrate that the snRNA gene promoter and enhancer elements, although very efficient in transcription of snRNAs, are unable to direct transcription of polyadenylated mRNAs. However, other sequences in the 5' flanking region of the human U1 gene can activate transcription of functional mRNA, with 5' ends upstream of the normal transcription start site.
我们研究了人类U1 RNA基因5'侧翼区序列指导功能性mRNA合成的能力。当与氯霉素乙酰转移酶(CAT)编码区序列融合时,U1基因的上游序列能够在293细胞中刺激功能性CAT mRNA的合成,但在HeLa细胞中则不能。293细胞中大多数多聚腺苷酸化的CAT mRNA起源于上游U1序列中的隐蔽启动子,但几乎所有从+1位置(相对于U1基因启动子)起始的CAT特异性RNA都是非多聚腺苷酸化的;这证实了真正的U1基因启动子无法指导多聚A+ mRNA的高效合成。我们的结果表明,小核RNA基因启动子和增强子元件虽然在小核RNA转录中非常高效,但无法指导多聚腺苷酸化mRNA的转录。然而,人类U1基因5'侧翼区的其他序列可以激活功能性mRNA的转录,其5'端位于正常转录起始位点的上游。