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J Virol. 1973 May; 11(5): 682-693
Copyright © 1973 American Society for Microbiology. All Rights Reserved.

Preferred Site for Initiation of RNA Transcription by Escherichia coli RNA Polymerase Within the Simian Virus 40 DNA Segment of the Nondefective Adenovirus-Simian Virus 40 Hybrid Viruses Ad2⁺ND₁ and Ad2⁺ND₃

Departments of Medicine, Human Genetics, and Molecular Biophysics and Biochemistry, Yale University School of Medicine, New Haven, Connecticut 06851
Laboratory of Viral Diseases, National Institute of Allergy and Infectious Diseases, Bethesda, Maryland 20014

ABSTRACT

The DNA of simian virus 40 (SV40) was transcribed into RNA by Escherichia coli RNA polymerase at 18 to 24 C after synchronization of the initiation of RNA synthesis. After a brief synthetic period the RNA product contained relatively large amounts of sequences derived from a limited segment of SV40 DNA. The source for this pulse-labeled RNA was found to be a portion of the segment of SV40 DNA included within the nondefective adenovirus (Ad)-SV40 hybrid viruses, Ad2⁺ND₁ and Ad2⁺ND₃. After synthesis with [-³²P] ATP, Ad2⁺ND₁ and Ad2⁺ND₃ DNA transcripts contained an initial sequence missing from Ad2 transcripts. This sequence was identified as an initiation sequence for polymerase transcription of the SV40 DNA. Thus, there is a preferred site for initiation of in vitro transcription on the segment of SV40 DNA common to the nondefective Ad2⁺ND₁ and Ad2⁺ND₃ hybrid viruses.