RNA polymerase components in Semliki Forest virus-infected cells: synthesis from large precursors

Abstract

Two previously undescribed stable polypeptides (referred to as nsp 90 and nsp 63) appear in mammalian and avian cells infected with Semliki Forest virus. They are distinguishable from the virus structural proteins and their known precursors by their molecular weights and tryptic peptide maps, and are identical in size to two polypeptides found in purified preparations of virus-specific RNA polymerase. Data from pulse-chase experiments and from the use of inhibitors of proteolytic cleavage indicate that nsp 90 and nsp 63 are synthesized via a series of post-translational cleavages from three larger polypeptides, p200, p184 and p150. The labelling kinetics after synchronous initiation of protein synthesis are also consistent with the synthesis of nsp 90 and nsp 63 from a common initiation site, and show that nsp 63 is located close to this site. It is concluded that nsp 90 and nsp 63 are components of the virus-specific RNA polymerase, and are synthesized via a post-translational cleavage scheme entirely separate from that leading to the synthesis of the virus structural proteins.