Scarlato V, Greene J R, Geiduschek E P
Department of Biology, University of California, San Diego, La Jolla 92093.
Virology. 1991 Feb;180(2):716-28. doi: 10.1016/0042-6822(91)90085-p.
Phage SPO1 middle transcripts are known to fall into two classes, m and m1l. Class m1l transcripts continue to be made late in the viral infection, while the synthesis of class m transcripts ceases soon after the onset of replication and late transcription. The experiments that are reported here deal with the regulatory nature of this diversity. The accumulation of transcripts associated with eight middle promoters was analyzed by S1 nuclease mapping. DNA sequence surrounding these middle promoters was determined or redetermined, and the stability of RNA associated with most of these promoters was also analyzed. Class m1l transcription was shown to be associated with SPO1 middle promoters that remain active at late stages of viral development, when middle promoters of class m are repressed. The consensus sequences of class m and m1l middle promoters were found to be indistinguishable and the search for sequences consensual with late promoters yielded only divergent candidates. No other consensus sequence that is specific and exclusive to either class of middle promoters was detected within a hundred base pairs upstream or downstream of these promoters. Considerable variations in the stabilities of SPO1 middle transcripts were found. Two promoters that are only 71 base pairs apart yielded transcripts that had substantially different stabilities. The 5'-flanking segment of the transcript associated with the upstream promoter apparently conferred a high degree of stability on this RNA.
已知噬菌体SPO1中期转录本分为两类,即m类和m1l类。m1l类转录本在病毒感染后期仍持续产生,而m类转录本的合成在复制和晚期转录开始后不久就停止了。本文报道的实验探讨了这种多样性的调控本质。通过S1核酸酶图谱分析了与八个中期启动子相关的转录本积累情况。确定或重新确定了这些中期启动子周围的DNA序列,还分析了与这些启动子中大多数相关的RNA的稳定性。结果表明,m1l类转录与SPO1中期启动子相关,这些启动子在病毒发育后期仍保持活性,而此时m类中期启动子受到抑制。发现m类和m1l类中期启动子的共有序列无法区分,并且寻找与晚期启动子一致的序列仅得到不同的候选序列。在这些启动子上游或下游一百个碱基对内,未检测到任何一类中期启动子特有的其他共有序列。发现SPO1中期转录本的稳定性存在相当大的差异。两个相距仅71个碱基对的启动子产生的转录本稳定性有很大不同。与上游启动子相关的转录本的5'侧翼片段显然赋予了该RNA高度的稳定性。