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Unusual splice sites in the E1A–E1B cotranscripts synthesized in adenovirus type 40-infected A549 cells

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Summary

The adenovirus E1 DNA region consists of two transcription units, E1A and E1B. In this paper we report that the E1A–E1B cotranscripts containing sequences of both the E1A and E1B regions are synthesized during adenovirus type 40 (Ad40) infection of A549 cells. Cytoplasmic RNA was isolated from Ad40-infected A549 cells at 24, 72, and 100h post infection (p.i.). The complementary (c) DNA was synthesized by reverse transcription using an oligo-dT primer and then amplified by the polymerase chain reaction (PCR) using primers derived from the E1A and E1B regions. The cDNAs thus amplified were sequenced either directly or after cloning into bacteriophage M13 vectors. Analysis of cDNA indicated that the E1A–E1B cotranscripts are synthesized at 72h p.i., but not at 24 or 100h p.i. Nucleotide sequences of three cDNAs of the E1A–E1B cotranscripts indicated that the cotranscripts originate from the E1A promoter and lack sequences for both the E1A poly(A) site and E1B cap site. The splices create open reading frames for E1A–E1B fused polypeptides around the E1A–E1B junctions in these mRNAs. Most interestingly, the sequence analysis showed that the 5′ and 3′ splice junctions in the two E1A–E1B cotranscripts do not conform to the splice consensus GT-AG rule. Our results thus suggest that factor(s) which lead to unsual splicing in the E1 mRNAs are present in Ad40-infected A549 cells.

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Authors and Affiliations

  1. Cancer Research Institute, Sapporo Medical College, Sapporo

    S. Ishida, Y. Fujinaga & K. Fujinaga

  2. Meiji Institute of Health Science, Meiji Milk Products Co., Odawara, Japan

    N. Sakamoto & S. Hashimoto

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  1. S. Ishida
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  2. Y. Fujinaga
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  3. K. Fujinaga
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  4. N. Sakamoto
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  5. S. Hashimoto
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Ishida, S., Fujinaga, Y., Fujinaga, K. et al. Unusual splice sites in the E1A–E1B cotranscripts synthesized in adenovirus type 40-infected A549 cells. Archives of Virology 139, 389–402 (1994). https://doi.org/10.1007/BF01310800

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  • DOI: https://doi.org/10.1007/BF01310800

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