Differential stimulation by polyamines of phage DNA-directed in vitro synthesis of proteins

Abstract

The effect of polyamines on T7- and λrif d18 DNA-directed synthesis of proteins in an Escherichia coli cell-free system has been studied. When T7 DNA was used as a template, the degree of stimulation by spermidine of protein synthesis was larger with T7 RNA polymerase than with M r 42 K protein, while the synthesis of M r 13.5 K protein was not stimulated significantly by spermidine. The synthesis of T7 RNA polymerase was stimulated approx. 10-fold by 1 mM spermidine. When λrif d18 DNA was used as a template, the synthesis of ββ′ subunits of RNA polymerase was stimulated greatly by spermidine, while the synthesis of elongation factor Tu and ribosomal proteins was not stimulated significantly by spermidine. Spermidine stimulation of T7 DNA-directed synthesis of T7 RNA polymerase was at the level of both translation and transcription. The degree of stimulation by spermidine was greater at the level of translation. Putrescine stimulated the synthesis of T7 RNA polymerase and M r 42 K protein to a small degree at the level of translation.