An ancient player unmasked: T4 rI encodes a t-specific antiholin.
Phage T4 effects lysis by its holin T and its endolysin E. Lysis is inhibited (LIN) if the infected cell is subjected to secondary infections by T4 phage particles. The T4 rI gene is required for LIN in all hosts tested. Here, we show that a cloned rI gene can impose a T-specific LIN on T-mediated lysis in the context of the phage lambda infective cycle, in the absence of other T4 genes and without secondary infection by T4. Moreover, it is shown that the T holin accumulates in the membrane during LIN, forming SDS-resistant oligomers. We show by cross-linking experiments that a T-RI heterodimer is formed during LIN, demonstrating that RI belongs to the functional class of antiholins, such as the S107 protein of lambda, which heterodimerizes with its cognate holin, S105. Finally, we show that the addition of Ni(2+) ions to the medium can block lysis by a T protein hexahistidine-tagged at its C-terminus, suggesting that liganding of the periplasmic domain is sufficient to impose lysis inhibition. The results are discussed in terms of a model in which the LIN-inducing signal of the secondary infecting phage influences a conformational equilibrium assumed by RI in the periplasm.
Ramanculov
E
E
Department of Biochemistry and Biophysics, 2128 TAMU, Texas A and M University, College Station, TX 77843-2128, USA.
Young
R
R
D000483:Alleles; D000595:Amino Acid Sequence; D017122:Bacteriophage T4 / Q000187:drug effects / Q000235:genetics* / Q000502:physiology*; D010582:Bacteriophage lambda / Q000235:genetics; D015153:Blotting, Western; D003001:Cloning, Molecular; D003432:Cross-Linking Reagents; D004926:Escherichia coli / Q000821:virology*; D005557:Formaldehyde; D015967:Gene Expression Regulation, Viral; D005814:Genes, Viral / Q000235:genetics / Q000502:physiology*; D008242:Lysogeny / Q000187:drug effects / Q000235:genetics* / Q000502:physiology; D008969:Molecular Sequence Data; D009532:Nickel / Q000378:metabolism / Q000494:pharmacology; D015398:Signal Transduction; D013379:Substrate Specificity; D014764:Viral Proteins / Q000037:antagonists & inhibitors* / Q000737:chemistry / Q000235:genetics / Q000378:metabolism
D003432:Cross-Linking Reagents; D014764:Viral Proteins; C120332:holin t protein, Enterobacteria phage T4; D005557:Formaldehyde; D009532:Nickel