Abstract
The IR1-U2 region of Epstein-Barr virus DNA consists of multiple copies of a 3.1-kilobase (kb) repeat sequence, IR1, which maps to the left of a 3.3-kb unique region, U2. Although hybridizations of cytoplasmic polyadenylated RNA from latently infected cells to viral DNA indicate that the IR1-U2 region encodes a substantial fraction of the viral RNA in these cells, only a single low-abundance 3-kb cytoplasmic polyadenylated RNA has been identified on Northern blots. Further analysis of the cytoplasmic polyadenylated RNA encoded by the IR1-U2 region indicates that (i) the RNA is transcribed from left to right; (ii) there are only three copies of the 3-kb RNA per cell; and (iii) the RNA is spliced. The RNA hybridizes to possibly contiguous 0.56- and 1.3-kb U2 domains. These domains and part of IR1 hybridize to the 3-kb cytoplasmic RNA. DNA between IR1 and the 0.56-kb U2 domain does not hybridize to the 3-kb RNA. The CCAAT-34 nucleotide-TATAA sequence in IR1 may be part of the promotor for the 3-kb cytoplasmic polyadenylated RNA since (i) it enables left-to-right transcription of IR1 by a HeLa cell extract, and (ii) latently infected cells contain giant polyadenylated nuclear RNAs which differ in size by 3 kb, as would be expected if transcription initiates in any copy of IR1 and continues through the rightward remaining copies into U2.
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Selected References
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- Acheson N. H., Buetti E., Scherrer K., Weil R. Transcription of the polyoma virus genome: synthesis and cleavage of giant late polyoma-specific RNA. Proc Natl Acad Sci U S A. 1971 Sep;68(9):2231–2235. doi: 10.1073/pnas.68.9.2231. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Aviv H., Leder P. Purification of biologically active globin messenger RNA by chromatography on oligothymidylic acid-cellulose. Proc Natl Acad Sci U S A. 1972 Jun;69(6):1408–1412. doi: 10.1073/pnas.69.6.1408. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Bailey J. M., Davidson N. Methylmercury as a reversible denaturing agent for agarose gel electrophoresis. Anal Biochem. 1976 Jan;70(1):75–85. doi: 10.1016/s0003-2697(76)80049-8. [DOI] [PubMed] [Google Scholar]
- Berger S. L., Birkenmeier C. S. Inhibition of intractable nucleases with ribonucleoside--vanadyl complexes: isolation of messenger ribonucleic acid from resting lymphocytes. Biochemistry. 1979 Nov 13;18(23):5143–5149. doi: 10.1021/bi00590a018. [DOI] [PubMed] [Google Scholar]
- Berk A. J., Lee F., Harrison T., Williams J., Sharp P. A. Pre-early adenovirus 5 gene product regulates synthesis of early viral messenger RNAs. Cell. 1979 Aug;17(4):935–944. doi: 10.1016/0092-8674(79)90333-7. [DOI] [PubMed] [Google Scholar]
- Berk A. J., Sharp P. A. Sizing and mapping of early adenovirus mRNAs by gel electrophoresis of S1 endonuclease-digested hybrids. Cell. 1977 Nov;12(3):721–732. doi: 10.1016/0092-8674(77)90272-0. [DOI] [PubMed] [Google Scholar]
- Bornkamm G. W., Hudewentz J., Freese U. K., Zimber U. Deletion of the nontransforming Epstein-Barr virus strain P3HR-1 causes fusion of the large internal repeat to the DSL region. J Virol. 1982 Sep;43(3):952–968. doi: 10.1128/jvi.43.3.952-968.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Cheung A., Kieff E. Long internal direct repeat in Epstein-Barr virus DNA. J Virol. 1982 Oct;44(1):286–294. doi: 10.1128/jvi.44.1.286-294.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Dambaugh T. R., Kieff E. Identification and nucleotide sequences of two similar tandem direct repeats in Epstein-Barr virus DNA. J Virol. 1982 Dec;44(3):823–833. doi: 10.1128/jvi.44.3.823-833.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Dambaugh T., Beisel C., Hummel M., King W., Fennewald S., Cheung A., Heller M., Raab-Traub N., Kieff E. Epstein-Barr virus (B95-8) DNA VII: molecular cloning and detailed mapping. Proc Natl Acad Sci U S A. 1980 May;77(5):2999–3003. doi: 10.1073/pnas.77.5.2999. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Donaldson D. S., McNab A. R., Rovera G., Curtis P. J. Nuclear precursor molecules of the two beta-globin mRNAs in Friend erythroleukemia cells. J Biol Chem. 1982 Aug 10;257(15):8655–8660. [PubMed] [Google Scholar]
- Gerper P., Whang-Peng J., Monroe J. H. Transformation and chromosome changes induced by Epstein-Barr virus in normal human leukocyte cultures. Proc Natl Acad Sci U S A. 1969 Jul;63(3):740–747. doi: 10.1073/pnas.63.3.740. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Goodman H. M., MacDonald R. J. Cloning of hormone genes from a mixture of cDNA molecules. Methods Enzymol. 1979;68:75–90. doi: 10.1016/0076-6879(79)68007-2. [DOI] [PubMed] [Google Scholar]
- Hayashi K. A cloning vehicle suitable for strand separation. Gene. 1980 Oct;11(1-2):109–115. doi: 10.1016/0378-1119(80)90091-8. [DOI] [PubMed] [Google Scholar]
- Hayward S. D., Lazarowitz S. G., Hayward G. S. Organization of the Epstein-Barr virus DNA molecule. II. Fine mapping of the boundaries of the internal repeat cluster of B95-8 and identification of additional small tandem repeats adjacent to the HR-1 deletion. J Virol. 1982 Jul;43(1):201–212. doi: 10.1128/jvi.43.1.201-212.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Heller M., Dambaugh T., Kieff E. Epstein-Barr virus DNA. IX. Variation among viral DNAs from producer and nonproducer infected cells. J Virol. 1981 May;38(2):632–648. doi: 10.1128/jvi.38.2.632-648.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Heller M., van Santen V., Kieff E. Simple repeat sequence in Epstein-Barr virus DNA is transcribed in latent and productive infections. J Virol. 1982 Oct;44(1):311–320. doi: 10.1128/jvi.44.1.311-320.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Hummel M., Kieff E. Epstein-Barr virus RNA. VIII. Viral RNA in permissively infected B95-8 cells. J Virol. 1982 Jul;43(1):262–272. doi: 10.1128/jvi.43.1.262-272.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Kawai Y., Nonoyama M., Pagano J. S. Reassociation kinetics for Epstein-Barr virus DNA: nonhomology to mammalian DNA and homology of viral DNA in various diseases. J Virol. 1973 Nov;12(5):1006–1012. doi: 10.1128/jvi.12.5.1006-1012.1973. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Kieff E., Dambaugh T., Heller M., King W., Cheung A., van Santen V., Hummel M., Beisel C., Fennewald S., Hennessy K. The biology and chemistry of Epstein-Barr virus. J Infect Dis. 1982 Oct;146(4):506–517. doi: 10.1093/infdis/146.4.506. [DOI] [PubMed] [Google Scholar]
- King W., Dambaugh T., Heller M., Dowling J., Kieff E. Epstein-Barr virus DNA XII. A variable region of the Epstein-Barr virus genome is included in the P3HR-1 deletion. J Virol. 1982 Sep;43(3):979–986. doi: 10.1128/jvi.43.3.979-986.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
- King W., Thomas-Powell A. L., Raab-Traub N., Hawke M., Kieff E. Epstein-Barr virus RNA. V. Viral RNA in a restringently infected, growth-transformed cell line. J Virol. 1980 Nov;36(2):506–518. doi: 10.1128/jvi.36.2.506-518.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
- King W., Van Santen V., Kieff E. Epstein-Barr virus RNA. VI. Viral RNA in restringently and abortively infected Raji cells. J Virol. 1981 May;38(2):649–660. doi: 10.1128/jvi.38.2.649-660.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Legon S., Flavell A. J., Cowie A., Kamen R. Amplification in the leader sequence of late polyoma virus mRNAs. Cell. 1979 Feb;16(2):373–388. doi: 10.1016/0092-8674(79)90013-8. [DOI] [PubMed] [Google Scholar]
- Lehrach H., Diamond D., Wozney J. M., Boedtker H. RNA molecular weight determinations by gel electrophoresis under denaturing conditions, a critical reexamination. Biochemistry. 1977 Oct 18;16(21):4743–4751. doi: 10.1021/bi00640a033. [DOI] [PubMed] [Google Scholar]
- Lindahl T., Adams A., Bjursell G., Bornkamm G. W., Kaschka-Dierich C., Jehn U. Covalently closed circular duplex DNA of Epstein-Barr virus in a human lymphoid cell line. J Mol Biol. 1976 Apr 15;102(3):511–530. doi: 10.1016/0022-2836(76)90331-4. [DOI] [PubMed] [Google Scholar]
- Manley J. L., Fire A., Cano A., Sharp P. A., Gefter M. L. DNA-dependent transcription of adenovirus genes in a soluble whole-cell extract. Proc Natl Acad Sci U S A. 1980 Jul;77(7):3855–3859. doi: 10.1073/pnas.77.7.3855. [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]
- McDonell M. W., Simon M. N., Studier F. W. Analysis of restriction fragments of T7 DNA and determination of molecular weights by electrophoresis in neutral and alkaline gels. J Mol Biol. 1977 Feb 15;110(1):119–146. doi: 10.1016/s0022-2836(77)80102-2. [DOI] [PubMed] [Google Scholar]
- Miller G., Robinson J., Heston L., Lipman M. Differences between laboratory strains of Epstein-Barr virus based on immortalization, abortive infection, and interference. Proc Natl Acad Sci U S A. 1974 Oct;71(10):4006–4010. doi: 10.1073/pnas.71.10.4006. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Nonoyama M., Pagano J. S. Separation of Epstein-Barr virus DNA from large chromosomal DNA in non-virus-producing cells. Nat New Biol. 1972 Aug 9;238(84):169–171. doi: 10.1038/newbio238169a0. [DOI] [PubMed] [Google Scholar]
- Orellana T., Kieff E. Epstein-barr virus-specific RNA. II. Analysis of polyadenylated viral RNA in restringent, abortive, and prooductive infections. J Virol. 1977 May;22(2):321–330. doi: 10.1128/jvi.22.2.321-330.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Pope J. H., Horne M. K., Scott W. Transformation of foetal human keukocytes in vitro by filtrates of a human leukaemic cell line containing herpes-like virus. Int J Cancer. 1968 Nov 15;3(6):857–866. doi: 10.1002/ijc.2910030619. [DOI] [PubMed] [Google Scholar]
- Powell A. L., King W., Kieff E. Epstein-Barr virus-specific RNA. III. Mapping of DNA encoding viral RNA in restringent infection. J Virol. 1979 Jan;29(1):261–274. doi: 10.1128/jvi.29.1.261-274.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Pritchett R. F., Hayward S. D., Kieff E. D. DNA of Epstein-Barr virus. I. Comparative studies of the DNA of Epstein-Barr virus from HR-1 and B95-8 cells: size, structure, and relatedness. J Virol. 1975 Mar;15(3):556–559. doi: 10.1128/jvi.15.3.556-559.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Pritchett R., Pendersen M., Kieff E. Complexity of EBV homologous DNA in continous lymphoblastoid cell lines. Virology. 1976 Oct 1;74(1):227–231. [PubMed] [Google Scholar]
- Raab-Traub N., Pritchett R., Kieff E. DNA of Epstein-Barr virus. III. Identification of restriction enzyme fragments that contain DNA sequences which differ among strains of Epstein-Barr virus. J Virol. 1978 Aug;27(2):388–398. doi: 10.1128/jvi.27.2.388-398.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Ragona G., Ernberg I., Klein G. Induction and biological characterization of the Epstein-Barr virus (EBV) carried by the Jijoye lymphoma line. Virology. 1980 Mar;101(2):553–557. doi: 10.1016/0042-6822(80)90473-0. [DOI] [PubMed] [Google Scholar]
- Reedman B. M., Klein G. Cellular localization of an Epstein-Barr virus (EBV)-associated complement-fixing antigen in producer and non-producer lymphoblastoid cell lines. Int J Cancer. 1973 May;11(3):499–520. doi: 10.1002/ijc.2910110302. [DOI] [PubMed] [Google Scholar]
- Rigby P. W., Dieckmann M., Rhodes C., Berg P. Labeling deoxyribonucleic acid to high specific activity in vitro by nick translation with DNA polymerase I. J Mol Biol. 1977 Jun 15;113(1):237–251. doi: 10.1016/0022-2836(77)90052-3. [DOI] [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]
- Svedmyr E., Jondal M. Cytotoxic effector cells specific for B Cell lines transformed by Epstein-Barr virus are present in patients with infectious mononucleosis. Proc Natl Acad Sci U S A. 1975 Apr;72(4):1622–1626. doi: 10.1073/pnas.72.4.1622. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Thomas P. S. Hybridization of denatured RNA and small DNA fragments transferred to nitrocellulose. Proc Natl Acad Sci U S A. 1980 Sep;77(9):5201–5205. doi: 10.1073/pnas.77.9.5201. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Treisman R. Characterisation of polyoma late mRNA leader sequences by molecular cloning and DNA sequence analysis. Nucleic Acids Res. 1980 Nov 11;8(21):4867–4888. doi: 10.1093/nar/8.21.4867. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Wellauer P. K., Dawid I. B. Secondary structure maps of RNA: processing of HeLa ribosomal RNA. Proc Natl Acad Sci U S A. 1973 Oct;70(10):2827–2831. doi: 10.1073/pnas.70.10.2827. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Zur Hausen H., Schulte-Holthausen H. Presence of EB virus nucleic acid homology in a "virus-free" line of Burkitt tumour cells. Nature. 1970 Jul 18;227(5255):245–248. doi: 10.1038/227245a0. [DOI] [PubMed] [Google Scholar]
- van Santen V., Cheung A., Kieff E. Epstein-Barr virus RNA VII: size and direction of transcription of virus-specified cytoplasmic RNAs in a transformed cell line. Proc Natl Acad Sci U S A. 1981 Mar;78(3):1930–1934. doi: 10.1073/pnas.78.3.1930. [DOI] [PMC free article] [PubMed] [Google Scholar]