Phage coat protein reacted with preparations of multi-stranded RNA a t high protein inputs to yield complexes containing phage-like particles. RNA complexes containing predominantly double-stranded RNA (replicative form, "RF") were considerably less efficient in promoting particle formation than those rich in single strands (replicative intermediate, "RI"). Similarly, binding of coat protein a t low input multiplicities was less efficient with "RF" than with "RI" preparations.The process of reduplication of those bacteriophages which contain RNA as their genetic material, has been investigated extensively in vivo and in vitro.Both approaches have led to the isolation of phagespecific, multi-stranded (i. e., partially doublestranded) RNA complexes [ 1,2] which are considered to be replicative intermediates [ 3,4]. Recent experiments revealed that multi-stranded RNA can be obtained from polysome fractions prepared from extracts of phage-infected cells [ 5,6]. This suggested that such structures are derived from complexes which may not only be involved in the process of transcription but in that of translation as well. Since their appearance in polysome fractions is apparent during late stages of infeehion, i. e. during those stages when the phage message is strongly modulated to code predominantly for coat protein [ 71, it seemed conceivable that the proposed RNA-modulation complex [S] would be contained in multi-stranded RNA and that such complexes might be formed in vitro.Furthermore, phage particle formation may take place in association with replicating complexes. Thus it should be possible to demonstrate the assembly of phage-like particles with preparations of multi-stranded RNA and coat protein subunits.The results of this study show that both types of RNA-coat protein complexes can be obtained , partially double-stranded; replicative form ("RP"), multistranded RNA not precipitable with 1.5 M NaCl [3], predominantly double-stranded; modulation complex, phage RNA to which one or a few coat protein subunits are attached and which translates the coat protein cistron but not the replicase cistron; assembly complex, phage RNA t o which sufficient coat protein subunits are attached to yield a phage-like particle.Enzyme. Pancreatic RNase or ribonucleate pyrimidinenucleotido-2'-transferase (cyclizing) (EC 2.7.7.16).
2'with multi-stranded phage RNA. Parts of these results have been presented elsewhere [9].
METHODS
Bacteria and BacteriophagesSingle colony isolates of Escherichia coli 3000 and 5000 F+ that resulted in good lysis after infection with an input multiplicity of 3-5 R17 bacteriophages were used. Bacteria and phages were grown as described [10,11].