Gabig Magdalena, Obuchowski Michal, WeLgrzyn Alicja, Szalewska-Palasz Agnieszka, Thomas Mark S, WeLgrzyn Grzegorz
Laboratory of Molecular Genetics, Department of Molecular Biology, University of GdańskKładki 24, 80-822 GdańskPoland.
Institute of Biochemistry and Biophysics, Polish Academy of Sciences, Laboratory of Molecular Biology affiliated to the University of GdańskK&lstroke;adki 24, 80-822 GdańskPoland.
Microbiology (Reading). 1998 Aug;144 ( Pt 8):2217-2224. doi: 10.1099/00221287-144-8-2217.
Bacteriophage lambda is unable to lysogenize Escherichia coli hosts harbouring the rpoA341 mutation due to a drastic reduction in transcription from CII-activated lysogenic promoters (pE, pI and paQ). In addition, the level of early transcripts involved in the lytic pathway of lambda development is also decreased in this genetic background due to impaired N-dependent antitermination. Here, it is demonstrated that despite the reduced level of early lytic pL- and pR-derived transcripts, lytic growth of bacteriophage lambda is not affected in rich media. The level of the late lytic, pR-derived transcripts also remains unaffected by the rpoA341 mutation under these conditions. However, it was found that whilst there is no significant difference in the phage burst size in rpoA+ and rpoA341 hosts growing in rich media, phage lambda is not able to produce progeny in the rpoA341 mutant growing in minimal medium, in contrast to otherwise isogenic rpoA+ bacteria. Provision of an excess of the phage replication proteins O and P in trans or overproduction of the antitermination protein N restore the ability of phage lambda to produce progeny in the rpoA341 mutant under the latter conditions. These results suggest that in rich media phage lambda produces some early proteins in excess of that needed for its effective propagation and indicate that replication proteins may be limiting factors for phage lytic growth in poor media.
由于CII激活的溶原性启动子(pE、pI和paQ)的转录大幅减少,噬菌体λ无法使携带rpoA341突变的大肠杆菌宿主发生溶原化。此外,在这种遗传背景下,由于N依赖的抗终止作用受损,参与λ发育裂解途径的早期转录本水平也会降低。在此,我们证明,尽管早期裂解性pL和pR衍生转录本的水平有所降低,但噬菌体λ在丰富培养基中的裂解生长并未受到影响。在这些条件下,晚期裂解性pR衍生转录本的水平也不受rpoA341突变的影响。然而,我们发现,虽然在丰富培养基中生长的rpoA+和rpoA341宿主中噬菌体的爆发量没有显著差异,但与其他同基因rpoA+细菌不同,噬菌体λ在基本培养基中生长的rpoA341突变体中无法产生后代。在后者的条件下,通过反式提供过量的噬菌体复制蛋白O和P或过量表达抗终止蛋白N,可以恢复噬菌体λ在rpoA341突变体中产生后代的能力。这些结果表明,在丰富培养基中,噬菌体λ产生的一些早期蛋白超过了其有效繁殖所需的量,并表明复制蛋白可能是噬菌体在贫瘠培养基中裂解生长的限制因素。