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A map of cytoplasmic RNA transcripts from lytic adenovirus type 2, determined by electron microscopy of RNANA hybrids
Cell, Volume 11, Issue 4, 819-836, 1 August 1977

Louise T. Chow,James M. Roberts,James B. LewisandThomas R. Broker

Cold Spring Harbor Laboratory Cold Spring Harbor, New York 11724 USA


Summary
We have used electron microscopic RNA loop mapping to determine, to within 200 nucleotides, the chromosome coordinates of the 5 and 3 ends of adenovirus type 2 transcripts isolated from the cytoplasm of productively infected human KB cells. The major blocks of transcription and many of the individual transcripts have discrete lengths and well defined 5 and 3 termini. Early cytoplasmic RNA is transcribed from four separate regions of the adenovirus genome, in map intervals 1.311.1, 62.467.9, 78.686.1 and 91.596.8. The most frequent late RNA loop occurred between coordinates 51.9 and 62.2. A late transcript complementary to interval 86.391.5 formed a characteristic R loop with a long branch of displaced RNA from coordinate 89.891.5, possibly a result of secondary structure near coordinate 89.8. Other late cytoplasmic RNA loops were located between coordinates 11.1 and 14.9, and in the intervals from 3052 and 6883. Transcripts from several regions of the chromosome had variable lengths but common 5 or 3 termini. Few R loops were seen between map coordinates 15 and 30. Switches between template strands were observed at map positions 11.2, 62.4 and 91.5. Many of the R loops have been correlated with gene products mapped previously by translation of RNA complementary to different portions of the chromosome.

An amazing sequence arrangement at the 5 ends of adenovirus 2 messenger RNA
Cell, Volume 12, Issue 1, 1-8, 1 September 1977

Louise T. Chow,Richard E. Gelinas,Thomas R. BrokerandRichard J. Roberts

Cold Spring Harbor Laboratory Cold Spring Harbor, New York 11724 USA


Summary
The 5 terminal sequences of several adenovirus 2 (Ad2) mRNAs, isolated late in infection, are complementary to sequences within the Ad2 genome which are remote from the DNA from which the main coding sequence of each mRNA is transcribed. This has been observed by forming RNA displacement loops (R loops) between Ad2 DNA and unfractionated polysomal RNA from infected cells. The 5 terminal sequences of mRNAs in R loops, variously located between positions 36 and 92, form complex secondary hybrids with single-stranded DNA from restriction endonuclease fragments containing sequences to the left of position 36 on the Ad2 genome. The structures visualized in the electron microscope show that short sequences coded at map positions 16.6, 19.6 and 26.6 on the R strand are joined to form a leader sequence of 150200 nucleotides at the 5 end of many late mRNAs. A late mRNA which maps to the left of position 16.6 shows a different pattern of second site hybridization. It contains sequences from 4.96.0 linked directly to those from 9.610.9. These findings imply a new mechanism for the biosynthesis of Ad2 mRNA in mammalian cells.

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