ADENOVIRUS-TYPE-12 VA RNA .1. SYNTHESIS IN PRODUCTIVE INFECTION AND GENE-MAPPING

VA RNA synthesized late in adenovirus type 12 (Ad12)-infected cells has been analyzed by two-dimensional polyacrylamide gel electrophoresis and by a sequence of three different gel electrophoresis separations. Each of these procedures resolves two distinct VA RNA species in Ad2-infected cells; no minor species was detected in either nuclei or cytoplasm of Ad12-infected KB cells. VA RNA synthesized in vitro in isolated nuclei of KB cells 18 hr after Ad12 infection also showed only one distinct species upon electrophoretic analysis. VA RNA synthesis in isolated nuclei was resistant to α-amanitin at concentration of 1 μg/ml but was inhibited by a high concentration of α-amanitin (100 μg/ml), which indicates that Ad12 VA RNA is synthesized by RNA polymerase III. The gene for VA RNA was mapped on Ad12 genome by blot hybridization experiments. It is located around position 0.30 of the unit map as determined by hybridization to fragments G and H of the BamHI digest of Ad12 DNA. The T1 ribonuclease oligonucleotide fingerprint analysis of RNA selected by hybridization to fragments G and H of the BamHU digest of Ad12 DNA has so far failed to provide evidence for two species of VA RNA of distinctly different structure in this adenovirus serotype. © 1979.