The UL47 gene (b 60390-b 60338) located in the unique long region of the human cytomegalovirus (HCMV) AD169 strain genome was analyzed by RNA mapping. Northern blot analysis showed that the UL47 gene was expressed at late times after infection (72 h postinfection). The 9.7-kb transcript was expressed in the infected cells but not in phosphonoformate-treated cells at 72 hpi, indicating that the UL47 gene was only expressed at late times after infection. To map the 5′-end and 3′-end of UL47 transcripts, primer extension and RNase protection analysis were performed. Primer extension analysis revealed that the transcription initiation site of UL47 was located in 27 bp downstream (b 60323) of the TATA box motif. The sizes of UL47 ORF (approximately 2.9-kb) and UL48 ORF (approximately 6.7-kb) deduced from computer sequence analysis suggest that the expressed 9.7-kb transcript of UL47 uses the 3′-end polyadenylation signal of UL48. The result of RNase protection determined that the 3′-end of UL47 RNA utilized the 3′-end polyadenylation signal of UL48, which is located in HCMV genome b 70082.
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Britt, W. J. and Alford, C. A., Cytomegalovirus;In Fields Virology. Fields, B. N. (Ed.). Lippincott-Raven, Philadelphia, 3rd Ed., Vol. 2, pp. 2493–2523, 1996.
Chang, C. P., Malone, C. L. and Stinski, M. F., A human cytomegalovirus early gene has three inducible promoters that are regulated differentially at various times after infection.J. Virol., 63, 281–290 (1989).
Chee, M. S., Bankier, A. T., Beck, S., Bohni, R., Brown, C. M., Cerny, R., Horsnell, T., Hutchison, C. A., Kouzarides, T., Martignetti, J. A., Preddie, E., Satchwell, S. C., Tomlinson, P., Weston, K. and Barrell, B. G., Analysis of the protein-coding content of the sequence of human cytomegalovirus strain AD169.Curr. Top. Microbiol. Immunol., 154, 125–169 (1990).
Davis, M. G. and Huang, E. S., Nucleotide sequence of a human cytomegalovirus DNA fragment encoding a 67-kilodalton phosphorylated viral protein.J. Virol., 56, 7–11 (1985).
DeMarchi, J. M., Human cytomegalovirus DNA: restriction enzyme cleavage maps and map locations of immediate-early, early, and late RNAs.Virology, 114, 23–38 (1981).
DeMarchi, J. M., Schmidt, C. A. and Kaplan, A. S., Patterns of transcription of human cytomegalovirus in permissively infected cells.J. Virol., 35, 227–286 (1980).
Depto, A. S. and Stenberg, R. M., Functional analysis of the true late human cytomegalovirus pp28 upstream promoter:cis-acting elements and viraltrans-acting proteins necessary for promoter activation.J. Virol., 66, 3241–3246 (1992).
Frederick, M. A., Roger, B., Robert, E. K., David, D. M., Seidmon, J. G., John, A. S. and Kevin, S.,Current protocols in molecular biology. John Wiley & Sons, Inc., 1997.
Geballe, A. P., Spaete, R. R. and Mocarski, E. S., Acis-acting element within the 5′ leader of a cytomegalovirus beta transcript determines kinetic class.Cell, 46, 865–872 (1986).
Jones, T. R. and Muzithras, V. P., Fine mapping of transcripts expressed from the US6 gene family of human cytomegalovirus strain AD169.J. Virol., 65, 2024–2036 (1991).
Kim, H. J., Regulation of the human cytomegalovirus US11 gene expression.Mol. Cells, 6, 61–66 (1996).
Klucher, K. M., Rabert, D. K. and Spector, D. H., Sequences in the human cytomegalovirus 2.7 kilobase RNA promoter which mediate its relation as an early gene.J. Virol., 63, 5334–5343 (1989).
Kouzarides, T., Bankier, A. T., Satchwell, S. C., Preddy, E. and Barrell, B. G., An immediate early gene of human cytomegalovirus encodes a potential membrane glycoprotein.Virology, 165, 151–164 (1988).
Kouzarides, T., Bankier, A. T., Satchwell, S. C., Weston, K., Tomlinson, P. and Barrell, B. G., Large scale rearrangement of homologous regions in the genomes of HCMV and EBV.Virology, 157, 397–413 (1987a).
Kouzarides, T., Bankier, A. T., Satchwell, S. C., Weston, K., Tomlinson, P. and Barrell, B. G., Sequence and transcription analysis of the human cytomegalovirus DNA polymerase gene.J. Virol., 61, 125–133 (1987b).
Lahijani, R. S., Otteson, E. W., Adlish, J. D. and St. Jeor, S. C., Characterization of a human cytomegalovirus 1.6 kilobase late mRNA and identification of its putative protein product.J. Virol., 65, 373–381 (1991).
Leach, F.S. and Mocarski, E. S., Regulation of cytomegalovirus late-gene expression: differential use of three start sites in the transcriptional activation of ICP36 gene expression.J. Virol., 63, 1783–1791 (1989).
Martinez, J., Lahijani, R. S. and St. Jeor, S. C., Analysis of a region of the human cytomegalovirus (AD169) genome coding for a 25-kllodalton virion protein.J. Virol., 63, 233–241 (1989).
Martinez, J. and St. Jeor, S. C., Molecular cloning and analysis of three cDNA clones homologous to human cytomegalovirus RNAs present during late infection.J. Virol., 60, 531–538 (1986).
McDonough, S. H. and Spector, D. H., Transcription in human fibroblasts permissively infected by human cytomegalovirus strain AD169.Virology, 125, 31–46 (1983).
Meyer, H., Bankier, A. T., Landini, M. P., Brown, C. M., Barrell, B. G., Ruger, B. and Mach, M., Identification and procaryotic expression of the gene coding for the highly immunogenic 28-kilodalton structural phosphoprotein (pp28) of human cytomegalovirus.J. Virol., 62, 2243–2250 (1988).
Mocarski, JR. E. S., Cytomegaloviruses and Their Replication;In Fields Virology. Fields, B. N. (Ed.). Lippincott-Raven Philadelphia, 3rd Ed., Vol. 2, pp. 2447–2492, 1996.
Pande, H., Campo, K., Churchill, M. A., Clark, B. R. and Zaia, J. A., Genomic localization of the gene encoding a 32-kDa capsid protein of human cytomegalovirus.Virology, 167, 306–310 (1988).
Roizman, B., Carmichael, L. E., Deinhardt, F., de-The, G., Nahmias, A. J., Plowright, W., Rapp, F., Sheldrick, P., Takahashi, M. and Wolf, K., Herpesviridae. Definition, provisional nomenclature, and taxonomy. The Herpesvirus Study Group, the International Committee on Taxonomy of Viruses.Intervirology, 16, 201–217 (1981).
Roizman, B. and Sears, A. E., Herpes Simplex Viruses and Their Replication;In Fields Virology. Fields., B. N. (Ed.). Lippincott-Raven, Philadelphia, 3rd Ed., Vol. 2, pp. 2231–2295, 1996.
Sambrook, J., Fritisch, E. F. and Maniatis, T.,Molecular Cloning: A Laboratory Manual, 2nd Ed., Cold Spring Harbor Laboratory Press, Cold Spring Harbor, New York, 1989.
Sanger, F., Nicklen, S. and Coulson, A. R., DNA sequencing with chain-terminating inhibitors.Proc. Natl. Acad. Sci. USA, 74, 5463–5467 (1977).
Spector, D. H., Klucher, K. M., Rabert, D. K. and Wright, D. A., Human cytomegalovirus early gene expression.Curr. Top. Microbiol. Immunol., 154, 21–45 (1990).
Staprans, S. I. and Spector, D. H., 2.2-kilobase class of early transcripts encoded by cell-related sequences in human cytomegalovirus strain AD169.J. Virol., 57, 591–602 (1986).
Stenberg, R. M., Fortney, J., Barlow, S. W., Magrane, B. P., Nelson, J. A. and Ghazal, P., Promoter-specific trans activation and repression by human cytomegalovirus immediate-early proteins involves common and unique protein domains.J. Virol., 64, 1556–1565 (1990).
Stenberg, R. M., Witte, P. R. and Stinski, M. F., Multiple spliced and unspliced transcripts from human cytomegalovirus immediate-early region 2 and evidence for a common initiation site within immediate-early region 1.J. Virol., 56, 665–675 (1985).
Stinski, M. F., Sequence of protein synthesis in cells infected by human cytomegalovirus: early and late virus-induced polypeptides.J. Virol., 26, 686–701 (1978).
Tenney, D. J. and Colberg-Poley, A. M., Expression of the human cytomegalovirus UL36-38 immediate-early region during permissive infection.Virology, 182, 199–210 (1991).
Wathen, M. W. and Stinski, M. F., Temporal patterns of human cytomegalovirus transcription: mapping the viral RNAs synthesized at immediate early, early, and late times after infection.J. Virol., 41, 462–477 (1982).
Wathen, M. W., Thomsen, D. R. and Stinski, M. F., Temporal regulation of human cytomegalovirus transcription at immediate early and early times after infection.J. Virol., 38, 446–459 (1981).
Weston, K., An enhancer element in the short unique region of human cytomegalovirus regulates the production of a group of abundant immediate early transcripts.Virology, 162, 406–416 (1988).
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Hyun, J., Park, H., Kim, K. et al. Analysis of transcripts expressed from the UL47 gene of human cytomegalovirus. Arch Pharm Res 22, 542–548 (1999). https://doi.org/10.1007/BF02975323
- Human cytomegalovirus
- Northern blot
- Primer extension
- RNase protection