Rz/Rz1 lysis gene equivalents in phages of Gram-negative hosts.
Under usual laboratory conditions, lysis by bacteriophage lambda requires only the holin and endolysin genes, but not the Rz and Rz1 genes, of the lysis cassette. Defects in Rz or Rz1 block lysis only in the presence of high concentrations of divalent cations. The lambda Rz and Rz1 lysis genes are remarkable in that Rz1, encoding an outer membrane lipoprotein, is completely embedded in the +1 register within Rz, which itself encodes an integral inner membrane protein. While Rz and Rz1 equivalents have been identified in T7 and P2, most phages, including such well-studied classic phages as T4, P1, T1, Mu and SP6, lack annotated Rz/Rz1 equivalents. Here we report that a search strategy based primarily on gene arrangement and membrane localization signals rather than sequence similarity has revealed that Rz/Rz1 equivalents are nearly ubiquitous among phages of Gram-negative hosts, with 120 of 137 phages possessing genes that fit the search criteria. In the case of T4, a deletion of a non-overlapping gene pair pseT.2 and pseT.3 identified as Rz/Rz1 equivalents resulted in the same divalent cation-dependent lysis phenotype. Remarkably, in T1 and six other phages, Rz/Rz1 pairs were not found but a single gene encoding an outer membrane lipoprotein with a C-terminal transmembrane domain capable of integration into the inner membrane was identified. These proteins were named "spanins," since their protein products are predicted to span the periplasm providing a physical connection between the inner and outer membranes. The T1 spanin gene was shown to complement the lambda Rz-Rz1- lysis defect, indicating that spanins function as Rz/Rz1 equivalents. The widespread presence of Rz/Rz1 or their spanin equivalents in phages of Gram-negative hosts suggests a strong selective advantage and that their role in the ecology of these phages is greater than that inferred from the mild laboratory phenotype.
Summer
Elizabeth J
EJ
Department of Biochemistry and Biophysics, 2128 TAMU, Texas A and M University, College Station, TX 77843-2128, USA.
Berry
Joel
J
Tran
Tram Anh T
TA
Niu
Lili
L
Struck
Douglas K
DK
Young
Ry
R
D001435:Bacteriophages / Q000235:genetics*; D015321:Gene Rearrangement; D006090:Gram-Negative Bacteria / Q000821:virology*; D008565:Membrane Proteins; D009154:Mutation; D010641:Phenotype; D014764:Viral Proteins / Q000235:genetics*
D008565:Membrane Proteins; D014764:Viral Proteins; C082268:Rz protein, bacteriophage lambda