Functional and structural features of the holin HOL protein of the Lactobacillus plantarum phage phi gle: analysis in Escherichia coli system.
Lactobacillus plantarum phage phi gle has two consecutive cell lysis genes hol-lys (Oki et al., 1996b). In the present study, functional and structural properties of the hol protein (Hol) were characterized in Escherichia coli. Electron microscopic examinations showed that hol under plac in E. coli XL1-Blue injured the inner membrane to yield empty ghost cells with the bulk of the cell wall undisturbed. Northern blot analysis indicated that hol-lys genes under plac were co-transcribed, although the amount of hol transcript was larger than that of lys, ceasing via an apparently rho-independent terminator just downstream of hol. However, deletion and/or fusion experiments suggested that: (1) the N-terminal half of phi gle Hol composed of three putative transmembrane domains may be responsible for interaction with membrane; (2) the N-terminal end (five amino acids) seems nonessential; and (3) the C-terminal half containing charged amino acids appears to be involved in proper hol function. These results suggest that phi gle Hol is a member of the lambdoid holin family, but divergent in several properties from lambda holin.
Oki
M
M
Molecular Biology Group, Faculty of Engineering, Toyama University, Japan.
Kakikawa
M
M
Nakamura
S
S
Yamamura
E T
ET
Watanabe
K
K
Sasamoto
M
M
Taketo
A
A
Kodaira
K
K
D000595:Amino Acid Sequence; D001433:Bacteriolysis; D001435:Bacteriophages / Q000235:genetics*; D001483:Base Sequence; D002462:Cell Membrane / Q000648:ultrastructure; D004279:DNA, Viral / Q000737:chemistry; D004926:Escherichia coli / Q000821:virology*; D005814:Genes, Viral / Q000235:genetics; D007778:Lactobacillus / Q000821:virology*; D008565:Membrane Proteins / Q000235:genetics* / Q000502:physiology; D008969:Molecular Sequence Data; D009154:Mutation; D009690:Nucleic Acid Conformation; D012367:RNA, Viral / Q000032:analysis; D011993:Recombinant Fusion Proteins; D015183:Restriction Mapping; D014764:Viral Proteins / Q000235:genetics* / Q000502:physiology; D015678:Viral Structural Proteins / Q000235:genetics
D004279:DNA, Viral; D008565:Membrane Proteins; D012367:RNA, Viral; D011993:Recombinant Fusion Proteins; D014764:Viral Proteins; D015678:Viral Structural Proteins