Pyrimidine tract binding protein strongly stimulates in vitro encephalomyocarditis virus RNA translation at the level of preinitiation complex formation

Abstract

Cellular protein p57/58, now known to be identical to polypyrimidine tract binding protein (PTB), has earlier been shown to specifically bind to the internal ribosome entry sites (IRES) of encephalomyocarditis virus (EMCV) and some other picornaviral RNAs. To elucidate its relevance to the internal initiation, the effect of cloned purified PTB on EMCV IRES directed translation was studied in cytoplasmic extracts of Krebs-2 ascites carcinoma cells partially depleted of endogenous PTB. Addition of PTB to such extracts resulted in a strong stimulation of translation of a beta-glucuronidase (GUS) reporter cistron fused to the EMCV IRES, but had no effect on translation of capped mRNAs, such as beta-globin, and tobacco mosaic virus (TMV) RNAs. PTB was found to exert its effect at the level of 48S pre-initiation complex formation.