Briat J F, Bollag G, Kearney C A, Molineux I, Chamberlin M J
Department of Biochemistry, University of California, Berkeley 94720.
J Mol Biol. 1987 Nov 5;198(1):43-9. doi: 10.1016/0022-2836(87)90456-6.
The termination signal that limits transcription through the early region of bacteriophage T3 (T3Te) has been cloned and sequenced. The nucleotide sequence of T3Te is identical with that of T7Te, with the exception of a single G to U substitution in the 3' tail of the terminated transcript, and addition of an AC to the loop in the terminator stem-loop, enlarging the loop to six residues. Previous studies of the properties of T3Te have shown that this site is rho independent and is highly efficient for termination in vivo, but is used poorly in vitro during transcription with purified Escherichia coli RNA polymerase. In contrast, the equivalent site in bacteriophage T7 (T7Te) is an efficient termination signal both in vivo and in vitro. However, T3Te becomes an efficient termination site in vitro in the presence of preparations of tau factor. This factor also alters the sites of RNA chain termination found in vitro at T3Te. Transcripts formed in the presence of tau are several nucleotides shorter than those produced with RNA polymerase alone, and have 3' termini that are almost identical with transcripts found in vivo. These latter results are similar to our earlier findings with T7Te, and suggest that other rho independent terminators may act with transcription termination factors in vivo.
限制噬菌体T3早期区域转录的终止信号(T3Te)已被克隆并测序。T3Te的核苷酸序列与T7Te的相同,只是在终止转录本的3'尾部有一个G到U的单碱基替换,并且在终止子茎环的环中添加了一个AC,使环扩展到六个残基。先前对T3Te特性的研究表明,该位点不依赖rho因子,在体内是高效的终止位点,但在体外使用纯化的大肠杆菌RNA聚合酶进行转录时效果不佳。相比之下,噬菌体T7中的等效位点(T7Te)在体内和体外都是有效的终止信号。然而,在存在tau因子制剂的情况下,T3Te在体外成为一个有效的终止位点。该因子还改变了在体外T3Te处发现的RNA链终止位点。在tau因子存在下形成的转录本比仅用RNA聚合酶产生的转录本短几个核苷酸,并且其3'末端与体内发现的转录本几乎相同。后一个结果与我们早期对T7Te的发现相似,表明其他不依赖rho因子的终止子在体内可能与转录终止因子共同起作用。