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. 1989 Jun;63(6):2746–2757. doi: 10.1128/jvi.63.6.2746-2757.1989

Negative regulatory element associated with potentially functional promoter and enhancer elements in the long terminal repeats of endogenous murine leukemia virus-related proviral sequences.

L Y Ch'ang 1, W K Yang 1, F E Myer 1, D M Yang 1
PMCID: PMC250771  PMID: 2542587

Abstract

Three series of recombinant DNA clones were constructed, with the bacterial chloramphenicol acetyltransferase (CAT) gene as a quantitative indicator, to examine the activities of promoter and enhancer sequence elements in the 5' long terminal repeat (LTR) of murine leukemia virus (MuLV)-related proviral sequences isolated from the mouse genome. Transient CAT expression was determined in mouse NIH 3T3, human HT1080, and mink CCL64 cultured cells transfected with the LTR-CAT constructs. The 700-base-pair (bp) LTRs of three polytropic MuLV-related proviral clones and the 750-bp LTRs of four modified polytropic proviral clones, in complete structures either with or without the adjacent downstream sequences, all showed very little or negligible activities for CAT expression, while ecotropic MuLV LTRs were highly active. The MuLV-related LTRs were divided into three portions and examined separately. The 3' portion of the MuLV-related LTRs that contains the CCAAC and TATAA boxes was found to be a functional promoter, being about one-half to one-third as active as the corresponding portion of ecotropic MuLV LTRs. A MboI-Bg/II fragment, representing the distinct 190- to 200-bp inserted segment in the middle, was found to be a potential enhancer, especially when examined in combination with the simian virus 40 promoter in CCL64 cells. A PstI-MboI fragment of the 5' portion, which contains the protein-binding motifs of the enhancer segment as well as the upstream LTR sequences, showed moderate enhancer activities in CCL6 cells but was virtually inactive in NIH 3T3 cells and HT1080 cells; addition of this fragment to the ecotropic LTR-CAT constructs depressed CAT expression. Further analyses using chimeric LTR constructs located the presence of a strong negative regulatory element within the region containing the 5' portion of the enhancer and the immediate upstream sequences in the MuLV-related LTRs.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Bacheler L. T. Molecular clones of endogenous murine leukemia virus-related DNA sequences from Balb/c mice: characterization of integration sites. Virology. 1984 Oct 15;138(1):129–142. doi: 10.1016/0042-6822(84)90153-3. [DOI] [PubMed] [Google Scholar]
  2. Blair D. G., McClements W. L., Oskarsson M. K., Fischinger P. J., Vande Woude G. F. Biological activity of cloned Moloney sarcoma virus DNA: Terminally redundant sequences may enhance transformation efficiency. Proc Natl Acad Sci U S A. 1980 Jun;77(6):3504–3508. doi: 10.1073/pnas.77.6.3504. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Boone L. R., Glover P. L., Innes C. L., Niver L. A., Bondurant M. C., Yang W. K. Fv-1 N- and B-tropism-specific sequences in murine leukemia virus and related endogenous proviral genomes. J Virol. 1988 Aug;62(8):2644–2650. doi: 10.1128/jvi.62.8.2644-2650.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Boone L. R., Myer F. E., Yang D. M., Kiggans J. O., Koh C., Tennant R. W., Yang W. K. Analysis of recombinant DNA clones of the endogenous BALB/c murine leukemia virus WN1802N: variation in long terminal repeat length. J Virol. 1983 Jan;45(1):484–488. doi: 10.1128/jvi.45.1.484-488.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Boone L. R., Myer F. E., Yang D. M., Ou C. Y., Koh C. K., Roberson L. E., Tennant R. W., Yang W. K. Reversal of Fv-1 host range by in vitro restriction endonuclease fragment exchange between molecular clones of N-tropic and B-tropic murine leukemia virus genomes. J Virol. 1983 Oct;48(1):110–119. doi: 10.1128/jvi.48.1.110-119.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Brand A. H., Breeden L., Abraham J., Sternglanz R., Nasmyth K. Characterization of a "silencer" in yeast: a DNA sequence with properties opposite to those of a transcriptional enhancer. Cell. 1985 May;41(1):41–48. doi: 10.1016/0092-8674(85)90059-5. [DOI] [PubMed] [Google Scholar]
  7. Chan H. W., Bryan T., Moore J. L., Staal S. P., Rowe W. P., Martin M. A. Identification of ecotropic proviral sequences in inbred mouse strains with a cloned subgenomic DNA fragment. Proc Natl Acad Sci U S A. 1980 Oct;77(10):5779–5783. doi: 10.1073/pnas.77.10.5779. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Chatis P. A., Holland C. A., Silver J. E., Frederickson T. N., Hopkins N., Hartley J. W. A 3' end fragment encompassing the transcriptional enhancers of nondefective Friend virus confers erythroleukemogenicity on Moloney leukemia virus. J Virol. 1984 Oct;52(1):248–254. doi: 10.1128/jvi.52.1.248-254.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Chattopadhyay S. K., Lander M. R., Gupta S., Rands E., Lowy D. R. Origin of mink cytopathic focus-forming (MCF) viruses:comparison with ecotropic and xenotropic murine leukemia virus genomes. Virology. 1981 Sep;113(2):465–483. doi: 10.1016/0042-6822(81)90175-6. [DOI] [PubMed] [Google Scholar]
  10. Clark S. P., Kaufhold R., Chan A., Mak T. W. Comparison of the transcriptional properties of the Friend and Moloney retrovirus long terminal repeats: importance of tandem duplications and of the core enhancer sequence. Virology. 1985 Jul 30;144(2):481–494. doi: 10.1016/0042-6822(85)90288-0. [DOI] [PubMed] [Google Scholar]
  11. Clark S. P., Mak T. W. Complete nucleotide sequence of an infectious clone of Friend spleen focus-forming provirus: gp55 is an envelope fusion glycoprotein. Proc Natl Acad Sci U S A. 1983 Aug;80(16):5037–5041. doi: 10.1073/pnas.80.16.5037. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Colicelli J., Goff S. P. Isolation of a recombinant murine leukemia virus utilizing a new primer tRNA. J Virol. 1986 Jan;57(1):37–45. doi: 10.1128/jvi.57.1.37-45.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. DesGroseillers L., Rassart E., Jolicoeur P. Thymotropism of murine leukemia virus is conferred by its long terminal repeat. Proc Natl Acad Sci U S A. 1983 Jul;80(14):4203–4207. doi: 10.1073/pnas.80.14.4203. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Dhar R., McClements W. L., Enquist L. W., Vande Woude G. F. Nucleotide sequences of integrated Moloney sarcoma provirus long terminal repeats and their host and viral junctions. Proc Natl Acad Sci U S A. 1980 Jul;77(7):3937–3941. doi: 10.1073/pnas.77.7.3937. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Fujita T., Shibuya H., Hotta H., Yamanishi K., Taniguchi T. Interferon-beta gene regulation: tandemly repeated sequences of a synthetic 6 bp oligomer function as a virus-inducible enhancer. Cell. 1987 May 8;49(3):357–367. doi: 10.1016/0092-8674(87)90288-1. [DOI] [PubMed] [Google Scholar]
  16. Gonzalez F. J., Nebert D. W. Autoregulation plus upstream positive and negative control regions associated with transcriptional activation of the mouse P1(450) gene. Nucleic Acids Res. 1985 Oct 25;13(20):7269–7288. doi: 10.1093/nar/13.20.7269. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Goodbourn S., Burstein H., Maniatis T. The human beta-interferon gene enhancer is under negative control. Cell. 1986 May 23;45(4):601–610. doi: 10.1016/0092-8674(86)90292-8. [DOI] [PubMed] [Google Scholar]
  18. Gorman C. M., Moffat L. F., Howard B. H. Recombinant genomes which express chloramphenicol acetyltransferase in mammalian cells. Mol Cell Biol. 1982 Sep;2(9):1044–1051. doi: 10.1128/mcb.2.9.1044. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Gorman C. M., Rigby P. W., Lane D. P. Negative regulation of viral enhancers in undifferentiated embryonic stem cells. Cell. 1985 Sep;42(2):519–526. doi: 10.1016/0092-8674(85)90109-6. [DOI] [PubMed] [Google Scholar]
  20. Hartley J. W., Wolford N. K., Old L. J., Rowe W. P. A new class of murine leukemia virus associated with development of spontaneous lymphomas. Proc Natl Acad Sci U S A. 1977 Feb;74(2):789–792. doi: 10.1073/pnas.74.2.789. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Herr W. Nucleotide sequence of AKV murine leukemia virus. J Virol. 1984 Feb;49(2):471–478. doi: 10.1128/jvi.49.2.471-478.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Hoggan M. D., O'Neill R. R., Kozak C. A. Nonecotropic murine leukemia viruses in BALB/c and NFS/N mice: characterization of the BALB/c Bxv-1 provirus and the single NFS endogenous xenotrope. J Virol. 1986 Dec;60(3):980–986. doi: 10.1128/jvi.60.3.980-986.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Holland C. A., Hartley J. W., Rowe W. P., Hopkins N. At least four viral genes contribute to the leukemogenicity of murine retrovirus MCF 247 in AKR mice. J Virol. 1985 Jan;53(1):158–165. doi: 10.1128/jvi.53.1.158-165.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Jolly D. J., Esty A. C., Subramani S., Friedmann T., Verma I. M. Elements in the long terminal repeat of murine retroviruses enhance stable transformation by thymidine kinase gene. Nucleic Acids Res. 1983 Mar 25;11(6):1855–1872. doi: 10.1093/nar/11.6.1855. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Joyner A., Yamamoto Y., Bernstein A. Retrovirus long terminal repeats activate expression of coding sequences for the herpes simplex virus thymidine kinase gene. Proc Natl Acad Sci U S A. 1982 Mar;79(5):1573–1577. doi: 10.1073/pnas.79.5.1573. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Khan A. S., Laigret F., Rodi C. P. Expression of mink cell focus-forming murine leukemia virus-related transcripts in AKR mice. J Virol. 1987 Mar;61(3):876–882. doi: 10.1128/jvi.61.3.876-882.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Khan A. S., Martin M. A. Endogenous murine leukemia proviral long terminal repeats contain a unique 190-base-pair insert. Proc Natl Acad Sci U S A. 1983 May;80(9):2699–2703. doi: 10.1073/pnas.80.9.2699. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Khan A. S., Rowe W. P., Martin M. A. Cloning of endogenous murine leukemia virus-related sequences from chromosomal DNA of BALB/c and AKR/J mice: identification of an env progenitor of AKR-247 mink cell focus-forming proviral DNA. J Virol. 1982 Nov;44(2):625–636. doi: 10.1128/jvi.44.2.625-636.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Khoury G., Gruss P. Enhancer elements. Cell. 1983 Jun;33(2):313–314. doi: 10.1016/0092-8674(83)90410-5. [DOI] [PubMed] [Google Scholar]
  30. Kozak C. A. Retroviruses as chromosomal genes in the mouse. Adv Cancer Res. 1985;44:295–336. doi: 10.1016/s0065-230x(08)60030-5. [DOI] [PubMed] [Google Scholar]
  31. Kuemmerle N. B., Ch'ang L. Y., Koh C. K., Boone L. R., Yang W. K. Characterization of two solitary long terminal repeats of murine leukemia virus type that are conserved in the chromosome of laboratory inbred mouse strains. Virology. 1987 Oct;160(2):379–388. doi: 10.1016/0042-6822(87)90009-2. [DOI] [PubMed] [Google Scholar]
  32. Kuhl D., de la Fuente J., Chaturvedi M., Parimoo S., Ryals J., Meyer F., Weissmann C. Reversible silencing of enhancers by sequences derived from the human IFN-alpha promoter. Cell. 1987 Sep 25;50(7):1057–1069. doi: 10.1016/0092-8674(87)90172-3. [DOI] [PubMed] [Google Scholar]
  33. Köhrer K., Grummt I., Horak I. Functional RNA polymerase II promoters in solitary retroviral long terminal repeats (LTR-IS elements). Nucleic Acids Res. 1985 Apr 11;13(7):2631–2645. doi: 10.1093/nar/13.7.2631. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Laimins L. A., Gruss P., Pozzatti R., Khoury G. Characterization of enhancer elements in the long terminal repeat of Moloney murine sarcoma virus. J Virol. 1984 Jan;49(1):183–189. doi: 10.1128/jvi.49.1.183-189.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Laimins L., Holmgren-König M., Khoury G. Transcriptional "silencer" element in rat repetitive sequences associated with the rat insulin 1 gene locus. Proc Natl Acad Sci U S A. 1986 May;83(10):3151–3155. doi: 10.1073/pnas.83.10.3151. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Lenz J., Celander D., Crowther R. L., Patarca R., Perkins D. W., Haseltine W. A. Determination of the leukaemogenicity of a murine retrovirus by sequences within the long terminal repeat. 1984 Mar 29-Apr 4Nature. 308(5958):467–470. doi: 10.1038/308467a0. [DOI] [PubMed] [Google Scholar]
  37. Levinson B., Khoury G., Vande Woude G., Gruss P. Activation of SV40 genome by 72-base pair tandem repeats of Moloney sarcoma virus. Nature. 1982 Feb 18;295(5850):568–572. doi: 10.1038/295568a0. [DOI] [PubMed] [Google Scholar]
  38. Levy D. E., Lerner R. A., Wilson M. C. Normal expression of polymorphic endogenous retroviral RNA containing segments identical to mink cell focus-forming virus. J Virol. 1985 Dec;56(3):691–700. doi: 10.1128/jvi.56.3.691-700.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  39. Levy D. E., Lerner R. A., Wilson M. C. The Gv-1 locus coordinately regulates the expression of multiple endogenous murine retroviruses. Cell. 1985 May;41(1):289–299. doi: 10.1016/0092-8674(85)90082-0. [DOI] [PubMed] [Google Scholar]
  40. Levy D. E., McKinnon R. D., Brolaski M. N., Gautsch J. W., Wilson M. C. The 3' long terminal repeat of a transcribed yet defective endogenous retroviral sequence is a competent promoter of transcription. J Virol. 1987 Apr;61(4):1261–1265. doi: 10.1128/jvi.61.4.1261-1265.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  41. Liou R. S., Boone L. R., Kiggans J. O., Yang D. M., Wang T. W., Tennant R. W., Yang W. K. Molecular cloning and analysis of the endogenous retrovirus chemically induced from RFM/Un mouse cell cultures. J Virol. 1983 Apr;46(1):288–292. doi: 10.1128/jvi.46.1.288-292.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  42. Loh T. P., Sievert L. L., Scott R. W. Proviral sequences that restrict retroviral expression in mouse embryonal carcinoma cells. Mol Cell Biol. 1987 Oct;7(10):3775–3784. doi: 10.1128/mcb.7.10.3775. [DOI] [PMC free article] [PubMed] [Google Scholar]
  43. Morley K. L., Toohey M. G., Peterson D. O. Transcriptional repression of a hormone-responsive promoter. Nucleic Acids Res. 1987 Sep 11;15(17):6973–6989. doi: 10.1093/nar/15.17.6973. [DOI] [PMC free article] [PubMed] [Google Scholar]
  44. Nikbakht K. N., Boone L. R., Glover P. L., Myer F. E., Yang W. K. Characterization of a molecular clone of RFM/Un mouse chromosomal DNA that contains a full-length endogenous murine leukaemia virus-related proviral genome. J Gen Virol. 1987 Mar;68(Pt 3):683–693. doi: 10.1099/0022-1317-68-3-683. [DOI] [PubMed] [Google Scholar]
  45. Nikbakht K. N., Ou C. Y., Boone L. R., Glover P. L., Yang W. K. Nucleotide sequence analysis of endogenous murine leukemia virus-related proviral clones reveals primer-binding sites for glutamine tRNA. J Virol. 1985 Jun;54(3):889–893. doi: 10.1128/jvi.54.3.889-893.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  46. Nir U., Walker M. D., Rutter W. J. Regulation of rat insulin 1 gene expression: evidence for negative regulation in nonpancreatic cells. Proc Natl Acad Sci U S A. 1986 May;83(10):3180–3184. doi: 10.1073/pnas.83.10.3180. [DOI] [PMC free article] [PubMed] [Google Scholar]
  47. O'Neill R. R., Khan A. S., Hoggan M. D., Hartley J. W., Martin M. A., Repaske R. Specific hybridization probes demonstrate fewer xenotropic than mink cell focus-forming murine leukemia virus env-related sequences in DNAs from inbred laboratory mice. J Virol. 1986 May;58(2):359–366. doi: 10.1128/jvi.58.2.359-366.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  48. Ostrowski M. C., Huang A. L., Kessel M., Wolford R. G., Hager G. L. Modulation of enhancer activity by the hormone responsive regulatory element from mouse mammary tumor virus. EMBO J. 1984 Aug;3(8):1891–1899. doi: 10.1002/j.1460-2075.1984.tb02064.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  49. Ou C. Y., Boone L. R., Yang W. K. A novel sequence segment and other nucleotide structural features in the long terminal repeat of a BALB/c mouse genomic leukemia virus-related DNA clone. Nucleic Acids Res. 1983 Aug 25;11(16):5603–5620. doi: 10.1093/nar/11.16.5603. [DOI] [PMC free article] [PubMed] [Google Scholar]
  50. Overhauser J., Fan H. Generation of glucocorticoid-responsive Moloney murine leukemia virus by insertion of regulatory sequences from murine mammary tumor virus into the long terminal repeat. J Virol. 1985 Apr;54(1):133–144. doi: 10.1128/jvi.54.1.133-144.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  51. Schmidt M., Glöggler K., Wirth T., Horak I. Evidence that a major class of mouse endogenous long terminal repeats (LTRs) resulted from recombination between exogenous retroviral LTRs and similar LTR-like elements (LTR-IS). Proc Natl Acad Sci U S A. 1984 Nov;81(21):6696–6700. doi: 10.1073/pnas.81.21.6696. [DOI] [PMC free article] [PubMed] [Google Scholar]
  52. Shinnick T. M., Lerner R. A., Sutcliffe J. G. Nucleotide sequence of Moloney murine leukaemia virus. Nature. 1981 Oct 15;293(5833):543–548. doi: 10.1038/293543a0. [DOI] [PubMed] [Google Scholar]
  53. Short M. K., Okenquist S. A., Lenz J. Correlation of leukemogenic potential of murine retroviruses with transcriptional tissue preference of the viral long terminal repeats. J Virol. 1987 Apr;61(4):1067–1072. doi: 10.1128/jvi.61.4.1067-1072.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  54. Speck N. A., Baltimore D. Six distinct nuclear factors interact with the 75-base-pair repeat of the Moloney murine leukemia virus enhancer. Mol Cell Biol. 1987 Mar;7(3):1101–1110. doi: 10.1128/mcb.7.3.1101. [DOI] [PMC free article] [PubMed] [Google Scholar]
  55. Steffen D. L., Mural R., Cowing D., Mielcarz J., Young J., Roblin R. Most of the murine leukemia virus sequences in the DNA of NIH/swiss mice consist of two closely related proviruses, each repeated several times. J Virol. 1982 Jul;43(1):127–135. doi: 10.1128/jvi.43.1.127-135.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  56. Stoye J. P., Coffin J. M. The four classes of endogenous murine leukemia virus: structural relationships and potential for recombination. J Virol. 1987 Sep;61(9):2659–2669. doi: 10.1128/jvi.61.9.2659-2669.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  57. Van Beveren C., van Straaten F., Galleshaw J. A., Verma I. M. Nucleotide sequence of the genome of a murine sarcoma virus. Cell. 1981 Nov;27(1 Pt 2):97–108. doi: 10.1016/0092-8674(81)90364-0. [DOI] [PubMed] [Google Scholar]
  58. Vogt M., Haggblom C., Swift S., Haas M. Envelope gene and long terminal repeat determine the different biological properties of Rauscher, Friend, and Moloney mink cell focus-inducing viruses. J Virol. 1985 Jul;55(1):184–192. doi: 10.1128/jvi.55.1.184-192.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  59. Wood T. G., McGeady M. L., Baroudy B. M., Blair D. G., Vande Woude G. F. Mouse c-mos oncogene activation is prevented by upstream sequences. Proc Natl Acad Sci U S A. 1984 Dec;81(24):7817–7821. doi: 10.1073/pnas.81.24.7817. [DOI] [PMC free article] [PubMed] [Google Scholar]
  60. Yoshimura F. K., Davison B., Chaffin K. Murine leukemia virus long terminal repeat sequences can enhance gene activity in a cell-type-specific manner. Mol Cell Biol. 1985 Oct;5(10):2832–2835. doi: 10.1128/mcb.5.10.2832. [DOI] [PMC free article] [PubMed] [Google Scholar]

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