Isolation of foot-and-mouth disease virus messenger RNA from membrane-bound polyribosomes and characterization of its 5′ and 3′ termini

Abstract

A membrane-bound fraction prepared from foot-and-mouth disease virus (FMDV)-infected baby hamster kidney cells included [ 3H]uridine-labeled 37 S viral messenger RNA (mRNA) and a 20 S RNA species that is partly ribonuclease resistant. The mRNA appeared to contain both a polyadenylic acid (poly (A)) and a polycytidylic acid (poly (C)) tract as demonstrated by its binding to oligo(dT)- and oligo(dG)-cellulose, respectively. Paper chromatography of an RNase T 2 digest of the [ 3H]adenosine-labeled poly(A) fragment from mRNA showed that the poly(A) is at the 3′ end of the mRNA and is approximately 45 nucleotides long. Digestion of 32P-labeled FMDV mRNA with RNase A and T 2 yielded pUp as the 5′ terminus; RNases A and T 2 digested FMD virion RNA lacked this structure.