Abstract
Four oligonucleotides, complementary to the adenovirus 2 (ad2) fiber mRNA were chemically synthesized and used as primers to study the 5' end of that messenger. The oligonucleotides which were 8, 10, 12, and 14 nucleotides long had a common 3'-terminal sequence, TAC, complementary to the initiating codon of the fiber mRNA. The 5'-32P-labeled oligonucleotides were hybridized to polyadenylated mRNA from ad2-infected cells and the primer was extended, using reverse transcriptase. When the resulting products were analyzed by polyacrylamide gel electrophoresis multiple distinct bands could be identified by autoradiography. The extension products were characterized by hybridization and sequence analysis. The most prominent band contained the well-known tripartite lead which is attached to many different late adenovirus mRNAs. In addition, four differently spliced leaders were characterized which, besides the three segments of the tripartite leader, contain one or two auxilliary segments. A kinetic analysis of the different leaders during lytic adenovirus infection shows no temporal variation.
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Selected References
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- Agarwal K. L., Brunstedt J., Noyes B. E. A general method for detection and characterization of an mRNA using an oligonucleotide probe. J Biol Chem. 1981 Jan 25;256(2):1023–1028. [PubMed] [Google Scholar]
- Akusjärvi G., Pettersson J. Sequence analysis of adenovirus DNA. IV. The genomic sequences encoding the common tripartite leader of late adenovirus messenger RNA. J Mol Biol. 1979 Oct 15;134(1):143–158. doi: 10.1016/0022-2836(79)90417-0. [DOI] [PubMed] [Google Scholar]
- Brawerman G., Mendecki J., Lee S. Y. A procedure for the isolation of mammalian messenger ribonucleic acid. Biochemistry. 1972 Feb 15;11(4):637–641. doi: 10.1021/bi00754a027. [DOI] [PubMed] [Google Scholar]
- Breathnach R., Benoist C., O'Hare K., Gannon F., Chambon P. Ovalbumin gene: evidence for a leader sequence in mRNA and DNA sequences at the exon-intron boundaries. Proc Natl Acad Sci U S A. 1978 Oct;75(10):4853–4857. doi: 10.1073/pnas.75.10.4853. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Chow L. T., Broker T. R., Lewis J. B. Complex splicing patterns of RNAs from the early regions of adenovirus-2. J Mol Biol. 1979 Oct 25;134(2):265–303. doi: 10.1016/0022-2836(79)90036-6. [DOI] [PubMed] [Google Scholar]
- Chow L. T., Broker T. R. The spliced structures of adenovirus 2 fiber message and the other late mRNAs. Cell. 1978 Oct;15(2):497–510. doi: 10.1016/0092-8674(78)90019-3. [DOI] [PubMed] [Google Scholar]
- Dorsett P. H., G8nsberg H. S. Characterization of type 5 adenovirus fiber protein. J Virol. 1975 Jan;15(1):208–216. doi: 10.1128/jvi.15.1.208-216.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Edge M. D., Green A. R., Heathcliffe G. R., Meacock P. A., Schuch W., Scanlon D. B., Atkinson T. C., Newton C. R., Markham A. F. Total synthesis of a human leukocyte interferon gene. Nature. 1981 Aug 20;292(5825):756–762. doi: 10.1038/292756a0. [DOI] [PubMed] [Google Scholar]
- Flint S. J., Sharp P. A. Adenovirus transcription. V. Quantitation of viral RNA sequences in adenovirus 2-infected and transformed cells. J Mol Biol. 1976 Sep 25;106(3):749–774. doi: 10.1016/0022-2836(76)90263-1. [DOI] [PubMed] [Google Scholar]
- Houghton M., Stewart A. G., Doel S. M., Emtage J. S., Eaton M. A., Smith J. C., Patel T. P., Lewis H. M., Porter A. G., Birch J. R. The amino-terminal sequence of human fibroblast interferon as deduced from reverse transcripts obtained using synthetic oligonucleotide primers. Nucleic Acids Res. 1980 May 10;8(9):1913–1931. doi: 10.1093/nar/8.9.1913. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Hérissé J., Courtois G., Galibert F. Nucleotide sequence of the EcoRI D fragment of adenovirus 2 genome. Nucleic Acids Res. 1980 May 24;8(10):2173–2192. doi: 10.1093/nar/8.10.2173. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Hérissé J., Galibert F. Nucleotide sequence of the EcoRI E fragment of adenovirus 2 genome. Nucleic Acids Res. 1981 Mar 11;9(5):1229–1240. doi: 10.1093/nar/9.5.1229. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Ishibashi M., Maizel J. V., Jr The polypeptides of adenovirus. VI. Early and late glycopolypeptides. Virology. 1974 Apr;58(2):345–361. doi: 10.1016/0042-6822(74)90070-1. [DOI] [PubMed] [Google Scholar]
- Klessig D. F., Chow L. T. Incomplete splicing and deficient accumulation of the fiber messenger RNA in monkey cells infected by human adenovirus type 2. J Mol Biol. 1980 May 15;139(2):221–242. doi: 10.1016/0022-2836(80)90306-x. [DOI] [PubMed] [Google Scholar]
- Levine A. J., Ginsberg H. S. Mechanism by which fiber antigen inhibits multiplication of type 5 adenovirus. J Virol. 1967 Aug;1(4):747–757. doi: 10.1128/jvi.1.4.747-757.1967. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Maxam A. M., Gilbert W. A new method for sequencing DNA. Proc Natl Acad Sci U S A. 1977 Feb;74(2):560–564. doi: 10.1073/pnas.74.2.560. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Maxam A. M., Gilbert W. Sequencing end-labeled DNA with base-specific chemical cleavages. Methods Enzymol. 1980;65(1):499–560. doi: 10.1016/s0076-6879(80)65059-9. [DOI] [PubMed] [Google Scholar]
- Mevarech M., Noyes B. E., Agarwal K. L. Detection of gastrin-specific mRNA using oligodeoxynucleotide probes of defined sequence. J Biol Chem. 1979 Aug 25;254(16):7472–7475. [PubMed] [Google Scholar]
- Persson H., Pettersson U., Mathews M. B. Synthesis of a structural adenovirus polypeptide in the absence of viral DNA replication. Virology. 1978 Oct 1;90(1):67–79. doi: 10.1016/0042-6822(78)90334-3. [DOI] [PubMed] [Google Scholar]
- Pettersson U., Philipson L., Höglund S. Structural proteins of adenoviruses. II. Purification and characterization of the adenovirus type 2 fiber antigen. Virology. 1968 Jun;35(2):204–215. doi: 10.1016/0042-6822(68)90261-4. [DOI] [PubMed] [Google Scholar]
- Philipson L., Lonberg-Holm K., Pettersson U. Virus-receptor interaction in an adenovirus system. J Virol. 1968 Oct;2(10):1064–1075. doi: 10.1128/jvi.2.10.1064-1075.1968. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Sood A. K., Pereira D., Weissman S. M. Isolation and partial nucleotide sequence of a cDNA clone for human histocompatibility antigen HLA-B by use of an oligodeoxynucleotide primer. Proc Natl Acad Sci U S A. 1981 Jan;78(1):616–620. doi: 10.1073/pnas.78.1.616. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Southern E. M. Detection of specific sequences among DNA fragments separated by gel electrophoresis. J Mol Biol. 1975 Nov 5;98(3):503–517. doi: 10.1016/s0022-2836(75)80083-0. [DOI] [PubMed] [Google Scholar]
- Sunquist B., Pettersson U., Thelander L., Philipson L. Structural proteins of adenoviruses. IX. Molecular weight and subunit composition of adenovirus type 2 fiber. Virology. 1973 Jan;51(1):252–256. doi: 10.1016/0042-6822(73)90389-9. [DOI] [PubMed] [Google Scholar]
- Zain S., Gingeras T. R., Bullock P., Wong G., Gelinas R. E. Determination and analysis of adenovirus-2 DNA sequences which may include signals for late messenger RNA processing. J Mol Biol. 1979 Dec 5;135(2):413–433. doi: 10.1016/0022-2836(79)90444-3. [DOI] [PubMed] [Google Scholar]
- Zain S., Sambrook J., Roberts R. J., Keller W., Fried M., Dunn A. R. Nucleotide sequence analysis of the leader segments in a cloned copy of adenovirus 2 fiber mRNA. Cell. 1979 Apr;16(4):851–861. doi: 10.1016/0092-8674(79)90100-4. [DOI] [PubMed] [Google Scholar]