Zajanckauskaite Aurelija, Truncaite Lidija, Strazdaite-Zieliene Zivile, Nivinskas Rimas
Department of Gene Engineering, Institute of Biochemistry, Vilnius, Lithuania.
Virology. 2008 Jun 5;375(2):342-53. doi: 10.1016/j.virol.2008.02.029. Epub 2008 Apr 18.
Sequence-specific endoribonuclease RegB of bacteriophage T4 cleaves early phage mRNAs and facilitates the transition between early and subsequent phases of T4 gene expression. The great majority of RegB targets have been identified in the intergenic regions of T4 transcripts, frequently in the Shine-Dalgarno sequences. Here we show that localization of RegB targets is not restricted to intergenic regions of mRNA. We detected 30 intragenic RegB sites in T4 transcripts that are differently susceptible to cleavage. Four RegB-processed mRNAs were previously shown to undergo further processing at so-called "secondary sites". We have found three additional transcripts carrying clear targets for both RegB and another endoribonuclease. We show that secondary cuts within RegB-processed T4 mRNAs are generated mainly by Escherichia coli RNase G, but that in some cases RNase E can recognize the same targets. Using plasmid-phage systems we demonstrate that T4 infection favours cleavage by the host endoribonucleases at these sites.
噬菌体T4的序列特异性核糖核酸内切酶RegB可切割早期噬菌体mRNA,并促进T4基因表达早期阶段与后续阶段之间的转换。RegB的绝大多数作用靶点已在T4转录本的基因间隔区中被鉴定出来,这些靶点经常位于Shine-Dalgarno序列中。在此我们表明,RegB作用靶点的定位并不局限于mRNA的基因间隔区。我们在T4转录本中检测到30个基因内RegB位点,这些位点对切割的敏感性各不相同。之前已表明,4种经RegB处理的mRNA会在所谓的“二级位点”进行进一步加工。我们发现另外3种转录本同时带有RegB和另一种核糖核酸内切酶的明确作用靶点。我们表明,经RegB处理的T4 mRNA中的二级切割主要由大肠杆菌核糖核酸酶G产生,但在某些情况下,核糖核酸酶E也能识别相同的靶点。利用质粒-噬菌体系统,我们证明T4感染有利于宿主核糖核酸内切酶在这些位点进行切割。