Extended function of plasmid partition genes: the Sop system of linear phage-plasmid N15 facilitates late gene expression.
The mitotic stability of the linear plasmid-prophage N15 of Escherichia coli depends on a partition system closely related to that of the F plasmid SopABC. The two Sop systems are distinguished mainly by the arrangement of their centromeric SopB-binding sites, clustered in F (sopC) and dispersed in N15 (IR1 to IR4). Because two of the N15 inverted repeat (IR) sites are located close to elements presumed (by analogy with phage lambda) to regulate late gene expression during the lytic growth of N15, we asked whether Sop partition functions play a role in this process. In N15, a putative Q antiterminator gene is located 6 kb upstream of the probable major late promoter and two intrinsic terminator-like sequences, in contrast to lambda, where the Q gene is adjacent to the late promoter. Northern hybridization and lacZ reporter activity confirmed the identity of the N15 late promoter (p52), demonstrated antiterminator activity of the Q analogue, and located terminator sequences between p52 and the first open reading frame. Following prophage induction, N15 mutated in IR2 (downstream from gene Q) or IR3 (upstream of p52) showed a pronounced delay in lysis relative to that for wild-type N15. Expression of ir3(-)-p52::lacZ during N15 wild-type lytic growth was strongly reduced relative to the equivalent ir3(+) fusion. The provision of Q protein and the IR2 and SopAB proteins in trans to ir3(+)-p52::lacZ increased expression beyond that seen in the absence of any one of these factors. These results indicate that the N15 Sop system has a dual role: partition and regulation of late gene transcription during lytic growth.
Ravin
Nikolai V
NV
Centre Bioengineering, Russian Academy of Sciences, 7-1 Prosp. 60 let Oktiabria, Moscow, 117312 Russia.
Rech
Jérôme
J
Lane
David
D
D001426:Bacterial Proteins / Q000378:metabolism*; D003090:Coliphages / Q000235:genetics*; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism / Q000821:virology*; D029968:Escherichia coli Proteins / Q000502:physiology*; D015967:Gene Expression Regulation, Viral / Q000502:physiology*; D005814:Genes, Viral / Q000235:genetics; D008242:Lysogeny / Q000235:genetics; D009876:Operon; D010957:Plasmids / Q000502:physiology*; D011533:Proviruses / Q000235:genetics; D014764:Viral Proteins / Q000378:metabolism / Q000502:physiology*
D001426:Bacterial Proteins; C099483:CB protein, N15 bacteriophage; D029968:Escherichia coli Proteins; C489223:SopB protein, E coli; D014764:Viral Proteins; C060971:SopA protein, Bacteria