1887

Abstract

SUMMARY

Bacteria infected with bacteriophage were disrupted with lysozyme and mild detergents and the intracellular phage-specific components resolved by sedimentation through neutral sucrose gradients. In pulse-chase experiments with [H]-thymidine most of the radioactivity initially appeared in a fast-sedimenting form of (fsf) which was very shear-sensitive and bound tightly to nitrocellulose. Label next appeared in a slower sedimenting form (ssf), then phage heads and finally virus particles. The ssf showed susceptibility to shear similar to that of from intact virus, and sedimented with it on neutral gradients. The ssf differed from virus by binding to nitrocellulose and showing a different sedimentation profile on alkaline-sucrose gradients. The pulse-labelled replicating was very heterogeneous in mol. wt. and appeared to contain many single-strand nicks which were extensively repaired while the was still in the fast-sedimenting form. The conversion sequence fsf ―→ ssf ―→ heads → phage was supported by the accumulation of components in non-permissive host bacteria infected with certain amber mutants of . One of these, . , could not form the phage-induced 5’ exonuclease and in these infections there was no conversion of replicating to ssf, and net synthesis stopped prematurely. It was concluded that maturation of virus involved mature virus-size pieces of of unusual structure as intermediates between replicating and phage heads. The process appeared to require functional induced exonuclease, but the role of this enzyme was unclear.

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1973-02-01
2022-01-23
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