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. 1987 May;61(5):1577–1585. doi: 10.1128/jvi.61.5.1577-1585.1987

Expression of the bovine leukemia virus X region in virus-infected cells.

N R Rice, S L Simek, G C Dubois, S D Showalter, R V Gilden, R M Stephens
PMCID: PMC254138  PMID: 3033284

Abstract

Bovine leukemia virus, like its closest relatives the human T-cell leukemia virus types I and II, contains a 1.8-kilobase X region between the env gene and the 3' long terminal repeat. In this communication, we report the detection and characterization of a subgenomic mRNA from which this X region is presumably translated. This mRNA was produced by a complex splicing mechanism which resulted in juxtaposition of the 5' end of the env gene and the two overlapping X-region open reading frames. Translation of this mRNA could yield at least two distinct proteins depending on which initiation codon is used. Detection of the protein encoded by the BLV X-region long open reading frame has been reported (N. Sagata, J. Tsuzuku-Kawamura, M. Nagayoshi-Aida, F. Shimizu, K.-I. Imagawa, and Y. Ikawa, Proc. Natl. Acad. Sci. USA 82:7879-7883, 1985). Using synthetic peptide antisera, we detected a protein encoded by the short open reading frame in virus-infected cells. The protein migrated in sodium dodecyl sulfate-polyacrylamide gels with an apparent molecular weight of 19,000. It is a nuclear phosphoprotein.

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  1. Aldovini A., De Rossi A., Feinberg M. B., Wong-Staal F., Franchini G. Molecular analysis of a deletion mutant provirus of type I human T-cell lymphotropic virus: evidence for a doubly spliced x-lor mRNA. Proc Natl Acad Sci U S A. 1986 Jan;83(1):38–42. doi: 10.1073/pnas.83.1.38. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. 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]
  3. 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]
  4. Bellini W. J., Englund G., Rozenblatt S., Arnheiter H., Richardson C. D. Measles virus P gene codes for two proteins. J Virol. 1985 Mar;53(3):908–919. doi: 10.1128/jvi.53.3.908-919.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Bender W., Chien Y. H., Chattopadhyay S., Vogt P. K., Gardner M. B., Davidson N. High-molecular-weight RNAs of AKR, NZB, and wild mouse viruses and avian reticuloendotheliosis virus all have similar dimer structures. J Virol. 1978 Mar;25(3):888–896. doi: 10.1128/jvi.25.3.888-896.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Bos J. L., Polder L. J., Bernards R., Schrier P. I., van den Elsen P. J., van der Eb A. J., van Ormondt H. The 2.2 kb E1b mRNA of human Ad12 and Ad5 codes for two tumor antigens starting at different AUG triplets. Cell. 1981 Nov;27(1 Pt 2):121–131. doi: 10.1016/0092-8674(81)90366-4. [DOI] [PubMed] [Google Scholar]
  7. Cann A. J., Rosenblatt J. D., Wachsman W., Shah N. P., Chen I. S. Identification of the gene responsible for human T-cell leukaemia virus transcriptional regulation. Nature. 1985 Dec 12;318(6046):571–574. doi: 10.1038/318571a0. [DOI] [PubMed] [Google Scholar]
  8. Capco D. G., Wan K. M., Penman S. The nuclear matrix: three-dimensional architecture and protein composition. Cell. 1982 Jul;29(3):847–858. doi: 10.1016/0092-8674(82)90446-9. [DOI] [PubMed] [Google Scholar]
  9. Copeland T. D., Morgan M. A., Oroszlan S. Complete amino acid sequence of the nucleic acid-binding protein of bovine leukemia virus. FEBS Lett. 1983 May 30;156(1):37–40. doi: 10.1016/0014-5793(83)80243-9. [DOI] [PubMed] [Google Scholar]
  10. Copeland T. D., Oroszlan S., Kalyanaraman V. S., Sarngadharan M. G., Gallo R. C. Complete amino acid sequence of human T-cell leukemia virus structural protein p15. FEBS Lett. 1983 Oct 17;162(2):390–395. doi: 10.1016/0014-5793(83)80793-5. [DOI] [PubMed] [Google Scholar]
  11. Curran J. A., Richardson C., Kolakofsky D. Ribosomal initiation at alternate AUGs on the Sendai virus P/C mRNA. J Virol. 1986 Feb;57(2):684–687. doi: 10.1128/jvi.57.2.684-687.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Deeley R. G., Gordon J. I., Burns A. T., Mullinix K. P., Binastein M., Goldberg R. F. Primary activation of the vitellogenin gene in the rooster. J Biol Chem. 1977 Nov 25;252(22):8310–8319. [PubMed] [Google Scholar]
  13. Deschamps J., Kettmann R., Burny A. Experiments with cloned complete tumor-derived bovine leukemia virus information prove that the virus is totally exogenous to its target animal species. J Virol. 1981 Nov;40(2):605–609. doi: 10.1128/jvi.40.2.605-609.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Ernst H., Shatkin A. J. Reovirus hemagglutinin mRNA codes for two polypeptides in overlapping reading frames. Proc Natl Acad Sci U S A. 1985 Jan;82(1):48–52. doi: 10.1073/pnas.82.1.48. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Felber B. K., Paskalis H., Kleinman-Ewing C., Wong-Staal F., Pavlakis G. N. The pX protein of HTLV-I is a transcriptional activator of its long terminal repeats. Science. 1985 Aug 16;229(4714):675–679. doi: 10.1126/science.2992082. [DOI] [PubMed] [Google Scholar]
  16. Giorgi C., Blumberg B. M., Kolakofsky D. Sendai virus contains overlapping genes expressed from a single mRNA. Cell. 1983 Dec;35(3 Pt 2):829–836. doi: 10.1016/0092-8674(83)90115-0. [DOI] [PubMed] [Google Scholar]
  17. Goldberg D. A. Isolation and partial characterization of the Drosophila alcohol dehydrogenase gene. Proc Natl Acad Sci U S A. 1980 Oct;77(10):5794–5798. doi: 10.1073/pnas.77.10.5794. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Gonda M. A., Rice N. R., Gilden R. V. Avian reticuloendotheliosis virus: characterization of the high-molecular-weight viral RNA in transforming and helper virus populations. J Virol. 1980 Jun;34(3):743–751. doi: 10.1128/jvi.34.3.743-751.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Graves D. C., Ferrer J. F. In vitro transmission and propagation of the bovine leukemia virus in monolayer cell cultures. Cancer Res. 1976 Nov;36(11 Pt 1):4152–4159. [PubMed] [Google Scholar]
  20. Grégoire D., Couez D., Deschamps J., Heuertz S., Hors-Cayla M. C., Szpirer J., Szpirer C., Burny A., Huez G., Kettmann R. Different bovine leukemia virus-induced tumors harbor the provirus in different chromosomes. J Virol. 1984 Apr;50(1):275–279. doi: 10.1128/jvi.50.1.275-279.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Kettmann R., Cleuter Y., Gregoire D., Burny A. Role of the 3' long open reading frame region of bovine leukemia virus in the maintenance of cell transformation. J Virol. 1985 Jun;54(3):899–901. doi: 10.1128/jvi.54.3.899-901.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Kettmann R., Cleuter Y., Mammerickx M., Meunier-Rotival M., Bernardi G., Burny A., Chantrenne H. Genomic integration of bovine leukemia provirus: comparison of persistent lymphocytosis with lymph node tumor form of enzootic. Proc Natl Acad Sci U S A. 1980 May;77(5):2577–2581. doi: 10.1073/pnas.77.5.2577. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Kettmann R., Deschamps J., Cleuter Y., Couez D., Burny A., Marbaix G. Leukemogenesis by bovine leukemia virus: proviral DNA integration and lack of RNA expression of viral long terminal repeat and 3' proximate cellular sequences. Proc Natl Acad Sci U S A. 1982 Apr;79(8):2465–2469. doi: 10.1073/pnas.79.8.2465. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Kiyokawa T., Seiki M., Iwashita S., Imagawa K., Shimizu F., Yoshida M. p27x-III and p21x-III, proteins encoded by the pX sequence of human T-cell leukemia virus type I. Proc Natl Acad Sci U S A. 1985 Dec;82(24):8359–8363. doi: 10.1073/pnas.82.24.8359. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Kozak M. Point mutations define a sequence flanking the AUG initiator codon that modulates translation by eukaryotic ribosomes. Cell. 1986 Jan 31;44(2):283–292. doi: 10.1016/0092-8674(86)90762-2. [DOI] [PubMed] [Google Scholar]
  26. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  27. Lee T. H., Coligan J. E., Sodroski J. G., Haseltine W. A., Salahuddin S. Z., Wong-Staal F., Gallo R. C., Essex M. Antigens encoded by the 3'-terminal region of human T-cell leukemia virus: evidence for a functional gene. Science. 1984 Oct 5;226(4670):57–61. doi: 10.1126/science.6089350. [DOI] [PubMed] [Google Scholar]
  28. Mamoun R. Z., Astier-Gin T., Kettmann R., Deschamps J., Rebeyrotte N., Guillemain B. J. The pX region of the bovine leukemia virus is transcribed as a 2.1-kilobase mRNA. J Virol. 1985 May;54(2):625–629. doi: 10.1128/jvi.54.2.625-629.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Mardon G., Varmus H. E. Frameshift and intragenic suppressor mutations in a Rous sarcoma provirus suggest src encodes two proteins. Cell. 1983 Mar;32(3):871–879. doi: 10.1016/0092-8674(83)90072-7. [DOI] [PubMed] [Google Scholar]
  30. Mount S. M. A catalogue of splice junction sequences. Nucleic Acids Res. 1982 Jan 22;10(2):459–472. doi: 10.1093/nar/10.2.459. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Nagashima K., Yoshida M., Seiki M. A single species of pX mRNA of human T-cell leukemia virus type I encodes trans-activator p40x and two other phosphoproteins. J Virol. 1986 Nov;60(2):394–399. doi: 10.1128/jvi.60.2.394-399.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Nigg E. A., Walter G., Singer S. J. On the nature of crossreactions observed with antibodies directed to defined epitopes. Proc Natl Acad Sci U S A. 1982 Oct;79(19):5939–5943. doi: 10.1073/pnas.79.19.5939. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Oroszlan S., Sarngadharan M. G., Copeland T. D., Kalyanaraman V. S., Gilden R. V., Gallo R. C. Primary structure analysis of the major internal protein p24 of human type C T-cell leukemia virus. Proc Natl Acad Sci U S A. 1982 Feb;79(4):1291–1294. doi: 10.1073/pnas.79.4.1291. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Rice N. R., Copeland T. D., Simek S., Oroszlan S., Gilden R. V. Detection and characterization of the protein encoded by the v-rel oncogene. Virology. 1986 Mar;149(2):217–229. doi: 10.1016/0042-6822(86)90123-6. [DOI] [PubMed] [Google Scholar]
  35. Rice N. R., Stephens R. M., Burny A., Gilden R. V. The gag and pol genes of bovine leukemia virus: nucleotide sequence and analysis. Virology. 1985 Apr 30;142(2):357–377. doi: 10.1016/0042-6822(85)90344-7. [DOI] [PubMed] [Google Scholar]
  36. Rice N. R., Stephens R. M., Couez D., Deschamps J., Kettmann R., Burny A., Gilden R. V. The nucleotide sequence of the env gene and post-env region of bovine leukemia virus. Virology. 1984 Oct 15;138(1):82–93. doi: 10.1016/0042-6822(84)90149-1. [DOI] [PubMed] [Google Scholar]
  37. 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]
  38. Rosen C. A., Sodroski J. G., Willems L., Kettmann R., Campbell K., Zaya R., Burny A., Haseltine W. A. The 3' region of bovine leukemia virus genome encodes a trans-activator protein. EMBO J. 1986 Oct;5(10):2585–2589. doi: 10.1002/j.1460-2075.1986.tb04538.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  39. Sagata N., Tsuzuku-Kawamura J., Nagayoshi-Aida M., Shimizu F., Imagawa K., Ikawa Y. Identification and some biochemical properties of the major XBL gene product of bovine leukemia virus. Proc Natl Acad Sci U S A. 1985 Dec;82(23):7879–7883. doi: 10.1073/pnas.82.23.7879. [DOI] [PMC free article] [PubMed] [Google Scholar]
  40. Sagata N., Yasunaga T., Ikawa Y. Two distinct polypeptides may be translated from a single spliced mRNA of the X genes of human T-cell leukemia and bovine leukemia viruses. FEBS Lett. 1985 Nov 11;192(1):37–42. doi: 10.1016/0014-5793(85)80038-7. [DOI] [PubMed] [Google Scholar]
  41. Sagata N., Yasunaga T., Tsuzuku-Kawamura J., Ohishi K., Ogawa Y., Ikawa Y. Complete nucleotide sequence of the genome of bovine leukemia virus: its evolutionary relationship to other retroviruses. Proc Natl Acad Sci U S A. 1985 Feb;82(3):677–681. doi: 10.1073/pnas.82.3.677. [DOI] [PMC free article] [PubMed] [Google Scholar]
  42. Schultz A. M., Copeland T. D., Oroszlan S. The envelope proteins of bovine leukemia virus: purification and sequence analysis. Virology. 1984 Jun;135(2):417–427. doi: 10.1016/0042-6822(84)90197-1. [DOI] [PubMed] [Google Scholar]
  43. Seiki M., Hattori S., Hirayama Y., Yoshida M. Human adult T-cell leukemia virus: complete nucleotide sequence of the provirus genome integrated in leukemia cell DNA. Proc Natl Acad Sci U S A. 1983 Jun;80(12):3618–3622. doi: 10.1073/pnas.80.12.3618. [DOI] [PMC free article] [PubMed] [Google Scholar]
  44. Seiki M., Hikikoshi A., Taniguchi T., Yoshida M. Expression of the pX gene of HTLV-I: general splicing mechanism in the HTLV family. Science. 1985 Jun 28;228(4707):1532–1534. doi: 10.1126/science.2990031. [DOI] [PubMed] [Google Scholar]
  45. Shaw M. W., Choppin P. W., Lamb R. A. A previously unrecognized influenza B virus glycoprotein from a bicistronic mRNA that also encodes the viral neuraminidase. Proc Natl Acad Sci U S A. 1983 Aug;80(16):4879–4883. doi: 10.1073/pnas.80.16.4879. [DOI] [PMC free article] [PubMed] [Google Scholar]
  46. Shimotohno K., Takahashi Y., Shimizu N., Gojobori T., Golde D. W., Chen I. S., Miwa M., Sugimura T. Complete nucleotide sequence of an infectious clone of human T-cell leukemia virus type II: an open reading frame for the protease gene. Proc Natl Acad Sci U S A. 1985 May;82(10):3101–3105. doi: 10.1073/pnas.82.10.3101. [DOI] [PMC free article] [PubMed] [Google Scholar]
  47. Slamon D. J., Shimotohno K., Cline M. J., Golde D. W., Chen I. S. Identification of the putative transforming protein of the human T-cell leukemia viruses HTLV-I and HTLV-II. Science. 1984 Oct 5;226(4670):61–65. doi: 10.1126/science.6089351. [DOI] [PubMed] [Google Scholar]
  48. Sodroski J., Rosen C., Goh W. C., Haseltine W. A transcriptional activator protein encoded by the x-lor region of the human T-cell leukemia virus. Science. 1985 Jun 21;228(4706):1430–1434. doi: 10.1126/science.2990028. [DOI] [PubMed] [Google Scholar]
  49. Staufenbiel M., Deppert W. Intermediate filament systems are collapsed onto the nuclear surface after isolation of nuclei from tissue culture cells. Exp Cell Res. 1982 Mar;138(1):207–214. doi: 10.1016/0014-4827(82)90107-0. [DOI] [PubMed] [Google Scholar]
  50. Tager H. S. Coupling of peptides to albumin with difluorodinitrobenzene. Anal Biochem. 1976 Apr;71(2):367–375. [PubMed] [Google Scholar]
  51. 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]
  52. Wachsman W., Golde D. W., Temple P. A., Orr E. C., Clark S. C., Chen I. S. HTLV x-gene product: requirement for the env methionine initiation codon. Science. 1985 Jun 28;228(4707):1534–1537. doi: 10.1126/science.2990032. [DOI] [PubMed] [Google Scholar]
  53. Yoshinaka Y., Oroszlan S. Bovine leukemia virus post-envelope gene coded protein: evidence for expression in natural infection. Biochem Biophys Res Commun. 1985 Aug 30;131(1):347–354. doi: 10.1016/0006-291x(85)91809-1. [DOI] [PubMed] [Google Scholar]

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