McLauchlan J, Gaffney D, Whitton J L, Clements J B
Nucleic Acids Res. 1985 Feb 25;13(4):1347-68. doi: 10.1093/nar/13.4.1347.
Our previous DNA sequence comparisons of 3' terminal portions from equivalent herpes simplex virus type 1 (HSV-1) and HSV-2 genes identified a conserved sequence (consensus YGTGTTYY; Y = pyrimidine) located approximately 30bp downstream from the AATAAA signal. We report here that this signal is located downstream from 67% of the mammalian mRNA 3' termini examined. Using constructions with the bacterial chloramphenicol acetyl transferase (CAT) gene linked to an HSV 'terminator' fragment, we show that deletions in the 'terminator' reduce CAT activities and the levels of CAT mRNA 3' termini. Specifically: (1) deletions of downstream sequences which extend up to the consensus YGTGTTYY signal reduce CAT levels to values 35% of those obtained with undeleted plasmids, (2) a deletion of a further 14bp, which removes the YGTGTTYY consensus but not the poly A site, reduces CAT activities to 1%-4%. The levels of CAT mRNA 3' termini reflect the reductions in CAT activities however, levels of mRNA 5' termini are unaffected by these deletions. The RNA produced in the absence of the YGTGTTYY signal is present in the cytoplasm although no CAT activity is detectable.
我们之前对单纯疱疹病毒1型(HSV-1)和HSV-2的等效基因3'末端部分进行的DNA序列比较,确定了一个保守序列(共有序列YGTGTTYY;Y =嘧啶),位于AATAAA信号下游约30bp处。我们在此报告,在所检测的哺乳动物mRNA 3'末端中,67%的末端下游都存在该信号。使用与HSV“终止子”片段相连的细菌氯霉素乙酰转移酶(CAT)基因构建体,我们发现“终止子”中的缺失会降低CAT活性以及CAT mRNA 3'末端的水平。具体而言:(1)向下游延伸至共有序列YGTGTTYY信号的序列缺失会使CAT水平降至未缺失质粒所得值的35%,(2)进一步缺失14bp,这会去除YGTGTTYY共有序列但不影响多聚腺苷酸化位点,使CAT活性降至1%-4%。CAT mRNA 3'末端的水平反映了CAT活性的降低,然而,mRNA 5'末端的水平不受这些缺失的影响。尽管检测不到CAT活性,但在没有YGTGTTYY信号的情况下产生的RNA存在于细胞质中。