The product of the bacteriophage lambda W gene: purification and properties.
Gene W is one of the 10 genes that control the morphogenesis of the bacteriophage lambda head. The morpho genesis of the phage lambda head proceeds through the synthesis of an intermediate assembly called the prohead. This is an empty shell into which the bacteriophage DNA is introduced--packaged--by the phage enzyme DNA terminase. The product of W (gpW) acts after DNA packaging, but before the addition of another phage product, gene product FII, and before the addition of tails. The role of gpW is unknown. The structure of N- and C-tagged gpW has been previously determined by nuclear magnetic resonance (NMR) spectroscopy. Here we report some of the properties of the native protein. The purification of gpW to homogeneity, overproduced by a plasmid derivative, is described. To obtain large amounts of the protein, the ribosome-binding site had to be modified, showing that inefficient translation of the message is the main mechanism limiting W gene expression. The molecular weight of the protein is in close agreement to the value predicted from the DNA sequence of the gene, which suggests that it is not post-transcriptionally modified. It behaves as a monomer in solution. Radioactively labeled gpW is incorporated into phage particles in in vitro complementation, showing that gpW is a structural protein. The stage at which gpW functions and other circumstantial evidence support the idea that six molecules of gpW polymerize on the connector before the incorporation of six molecules of gpFII and before the tail attaches.
Murialdo
Helios
H
Fundación Ciencia para la Vida and Millennium Institute for Fundamental and Applied Biology, Avenida Marathon 1943, Santiago, Chile. hml@bionova.cl
Xing
Xuekun
X
Tzamtzis
Dimitra
D
Haddad
Abraham
A
Gold
Marvin
M
D010582:Bacteriophage lambda / Q000235:genetics*; D002850:Chromatography, Gel; D004274:DNA, Recombinant; D004706:Endodeoxyribonucleases / Q000378:metabolism; D005814:Genes, Viral; D013058:Mass Spectrometry; D010957:Plasmids / Q000235:genetics; D013462:Sulfur Radioisotopes; D015678:Viral Structural Proteins / Q000096:biosynthesis* / Q000302:isolation & purification
D004274:DNA, Recombinant; D013462:Sulfur Radioisotopes; D015678:Viral Structural Proteins; C411883:protein W, bacteriophage lambda; D004706:Endodeoxyribonucleases; C030894:terminase