Proteolytic processing of phage lambda tail protein gpH: timing of the cleavage

Abstract

We describe a method for the rapid partial purification of intermediate structures of phage lambda tail assembly, using formaldehyde-fixed Escherichia coli cells to precipitate tail-related structures. The purification depends on the specific interaction between the E. coli lambda receptor protein and lambda tail protein gpJ. Protein compositions of tail assembly intermediates were analyzed to determine when in the assembly sequence the minor tail protein gpH is cleaved. gpH joins the tail precursor structure early in the pathway, during assembly of the initiator (a structure that becomes the tail tip). However, gpH is not cleaved until after initiator assembly is complete and after the tail shaft has polymerized onto the initiator. These results suggest that each gpH molecule is extended along the length of the tail. Our results also appear to eliminate an ambiguity in the tail assembly pathway determined by earlier experiments: we argue that gene G acts between genes H and M.