Wijffelman C, van de Putte P
Mol Gen Genet. 1974;135(4):327-37. doi: 10.1007/BF00271147.
It has previously been shown that the transcription of Mu is asymmetric and takes place on the heavy DNA strand (Bade, 1972; Wijffelman et al., 1974). The direction of transcription of Mu has now been determined by RNA-DNA hybridizations between purified Mu-RNA and the separated strands of lambda-Mu hybrid phages. The direction of transcription is from the c-gene (immunity gene) end of the heavy strand to the beta-end (immunity distal end) (Fig. 1). Thermo-inducible, defective Mu lysogens, in which the prophage is deleted from the beta-end, have a normal early transcription pattern, but the increase of RNA at later times is absent. A defective lysogen, which contains only the immunity gene c and the genes A and B, still has an early transcription pattern similar to that of the wild-type. Therefore, we conclude that the early RNA is transcribed from that region of the Mu genome. The early Mu-RNA synthesis is negatively regulated with a minimum rate of transcription at 9 minutes after induction. Before the onset of the late RNA synthesis, at about 22 minutes there is a rather long period in which the rate of Mu-RNA synthesis slowly increases. Using DNA strands of lambda-Mu hybrids which contain only that part of the Mu-DNA on which the early RNA synthesis takes place, we have determined that during the first half in the intermediate phase only early genes are transcribed. The amount of Mu-RNA synthesized by a Mu prophage carrying the X-mutation, which influences the excision of Mu, is greatly reduced. Negative regulation of early transcription occurs normally in this mutant.
先前已经表明,Mu的转录是不对称的,并且发生在重DNA链上(巴德,1972;维耶费尔曼等人,1974)。现在,通过纯化的Mu-RNA与λ-Mu杂交噬菌体的单链之间的RNA-DNA杂交,确定了Mu的转录方向。转录方向是从重链的c基因(免疫基因)端到β端(免疫远端)(图1)。热诱导的缺陷型Mu溶原菌,其中原噬菌体从β端缺失,具有正常的早期转录模式,但后期RNA的增加不存在。一种缺陷型溶原菌,仅包含免疫基因c以及基因A和B,仍然具有与野生型相似的早期转录模式。因此,我们得出结论,早期RNA是从Mu基因组的该区域转录而来的。早期Mu-RNA合成受到负调控,诱导后9分钟转录速率最低。在晚期RNA合成开始之前,大约在22分钟时有一段相当长的时间,Mu-RNA合成速率缓慢增加。使用仅包含Mu-DNA上早期RNA合成发生部分的λ-Mu杂交体的DNA链,我们确定在中间阶段的前半段仅转录早期基因。携带影响Mu切除的X突变的Mu原噬菌体合成的Mu-RNA量大大减少。在该突变体中,早期转录的负调控正常发生。