The pI and pXI assembly proteins serve separate and essential roles in filamentous phage assembly.
Three non-capsid, phage-encoded proteins, pI, pIV and pXI, are required for assembly of the filamentous bacteriophage at the envelope of Escherichia coli. pIV forms the outer membrane component of the assembly site, and pI and pXI are predicted to form the cytoplasmic membrane component. pXI is the result of an in-frame internal translational initiation event in gene I and is identical with the carboxyl-terminal third of pI in amino acid sequence, membrane localization and topology. The two proteins share a cytoplasmic domain predicted to be an amphipathic helix, a transmembrane domain, and a periplasmic domain. By mutating the initiation site for pXI, a phage was made that produced only pI and was shown to absolutely require functional plasmid-encoded pXI for growth. Further mutational analysis was done to examine the functional determinants of the amphipathic helix and periplasmic domains of the pI and pXI proteins. The results show that the amphipathic helix region is very important for pI function but not for pXI function. Mutational analysis of the periplasmic domains of pI and pXI implies that these domains also perform separate functions, and suggests that the interaction between pI and pIV in the periplasm is critical for assembly. The results are discussed with regard to the separate roles that the pI and pXI proteins play in the overall process of phage assembly.
Haigh
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NG
Department of Biochemistry, Duke University Medical Center, Durham, NC 27710, USA.
Webster
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RE
D000595:Amino Acid Sequence; D004926:Escherichia coli / Q000166:cytology / Q000821:virology; D005816:Genetic Complementation Test; D017912:Inovirus / Q000235:genetics / Q000254:growth & development*; D008969:Molecular Sequence Data; D009154:Mutation / Q000235:genetics; D019897:Periplasm / Q000378:metabolism / Q000821:virology; D010641:Phenotype; D017433:Protein Structure, Secondary; D017386:Sequence Homology, Amino Acid; D013329:Structure-Activity Relationship; D017361:Viral Nonstructural Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D019065:Virus Assembly
D017361:Viral Nonstructural Proteins; C402507:non-capsid protein pXI, Filamentous bacteriophage; C402506:pI protein, Filamentous phage phiLf; C119520:pIV protein, Enterobacteria phage f1