The high-resolution functional map of bacteriophage SPP1 portal protein.
An essential component in the assembly of nucleocapsids of tailed bacteriophages and of herpes viruses is the portal protein that is located at the unique vertex of the icosahedral capsid through which DNA movements occur. A library of mutations in the bacteriophage SPP1 portal protein (gp6) was generated by random mutagenesis of gene 6. Screening of the library allowed identification of 67 single amino acid substitutions that impair portal protein function. Most of the mutations cluster within stretches of a few amino acids in the gp6 carboxyl-terminus. The mutations were divided into five classes according to the step of virus assembly that they impair: (1) production of stable gp6; (2) interaction of gp6 with the minor capsid protein gp7; (3) incorporation of gp6 in the procapsid structure; (4) DNA packaging; and (5) sizing of the packaged DNA molecule. Most of the mutations fell in classes 3 and 4. This is the first high-resolution functional map of a portal protein, in which its function at different steps of viral assembly can be directly correlated with specific regions of its sequence. The work provides a framework for the understanding of central processes in the assembly of viruses that use specialized portals to govern entry and exit of DNA from the viral capsid.
Isidro
Anabela
A
Max-Plank Institut für Molekulare Genetik, Berlin, Germany.
Santos
Mário A
MA
Henriques
Adriano O
AO
Tavares
Paulo
P
D019943:Amino Acid Substitution; D017103:Bacillus Phages / Q000235:genetics / Q000502:physiology*; D001412:Bacillus subtilis / Q000821:virology; D004252:DNA Mutational Analysis; D042003:DNA Packaging; D004279:DNA, Viral / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism; D005814:Genes, Viral; D016296:Mutagenesis; D020125:Mutation, Missense; D016133:Polymerase Chain Reaction; D011485:Protein Binding; D017422:Sequence Analysis, DNA; D014764:Viral Proteins / Q000737:chemistry / Q000235:genetics* / Q000502:physiology*; D015678:Viral Structural Proteins / Q000235:genetics / Q000502:physiology; D019065:Virus Assembly / Q000235:genetics / Q000502:physiology
D004279:DNA, Viral; D014764:Viral Proteins; D015678:Viral Structural Proteins; C115880:portal protein, Bacteriophage SPP1