Analysis of Parvovirus mRNA by Sedimentation and Electrophoresis in Aqueous and Nonaqueous Solution

Abstract

Adenovirus-associated virus (AAV)-specific RNA present in the cytoplasm of cells coinfected with a helper adenovirus was analyzed by sucrose gradient sedimentation and gel electrophoresis. In aqueous conditions both gels or gradients revealed three AAV RNA components corresponding to 30 S , 27 S , and 20 S and having apparent molecular weights of 2.6 × 10 6 , 1.75 × 10 6 to 1.8 × 10 6 , and 0.9 × 10 6 to 1.0 × 10 6 , respectively. In nonaqueous, denaturing solvents only the 20 S AAV RNA species was observed. For this reason, and because they would be apparently significantly larger than a single AAV DNA strand, both the 30 S and 27 S species are believed to result from conformational or aggregation effects in the aqueous nondenaturing systems. It is concluded that only a single RNA molecule having a molecular weight of approximately 0.9 × 10 6 to 1.0 × 10 6 is synthesized by AAV.