Isolation of a transcriptive complex from Newcastle disease virions

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Abstract
An active transcriptive complex was isolated from purified virions of Newcastle disease virus. After disruption with Triton X-100 and high salt, soluble and particulate fractions were separated by density gradient centrifugation. The transcriptive complex, recovered at a density of 1.275 g/cm3, appeared as a nucleocapsid structure by EM. When analyzed by polyacrylamide gel electrophoresis, the nucleocapsids consisted of the nucleocapsid protein, a minor protein of 53,000 MW, and the large L protein. Nucleocapsids possessed less than 1% of the hemagglutinating and neuraminidase activities originally associated with virions. The active complex synthesized predominantly 1120S RNA in vitro, and approximately 1/4 of the RNA molecules contained polyadenylic acid segments. In the presence of S-adenosyl-L-methionine, the RNA molecules were capped and methylated at the 5'' termini. The transcriptive complex was also capable of methylating exogenous Escherichia coli RNA in the absence of viral RNA synthesis.