Abstract
The evolutionarily conserved protein domain, called the Krüppel-associated box (KRAB), present in the amino termini of a large number of Krüppel-type zinc finger proteins is a strong repressor domain. In order to develop novel strategies to control human immunodeficiency virus type 1 (HIV-1) gene expression, we constructed a series of expression vectors expressing the wild-type Tat or Tat transdominant negative mutants fused to the KRAB repressor domain. We found that the KRAB domain tethered to the transactivating response element is able to suppress both basal and Tat-mediated activity of HIV-1 long terminal repeat-driven gene expression. These results suggest that the KRAB repressor domain fused to the Tat transdominant negative mutants can be successfully employed to control HIV-1 gene expression.
Full Text
The Full Text of this article is available as a PDF (184.7 KB).
Selected References
These references are in PubMed. This may not be the complete list of references from this article.
- Balboni P. G., Bozzini R., Zucchini S., Marconi P. C., Grossi M. P., Caputo A., Manservigi R., Barbanti-Brodano G. Inhibition of human immunodeficiency virus reactivation from latency by a tat transdominant negative mutant. J Med Virol. 1993 Dec;41(4):289–295. doi: 10.1002/jmv.1890410406. [DOI] [PubMed] [Google Scholar]
- Baltimore D. Gene therapy. Intracellular immunization. Nature. 1988 Sep 29;335(6189):395–396. doi: 10.1038/335395a0. [DOI] [PubMed] [Google Scholar]
- Choo Y., Sánchez-García I., Klug A. In vivo repression by a site-specific DNA-binding protein designed against an oncogenic sequence. Nature. 1994 Dec 15;372(6507):642–645. doi: 10.1038/372642a0. [DOI] [PubMed] [Google Scholar]
- Eguchi Y., Itoh T., Tomizawa J. Antisense RNA. Annu Rev Biochem. 1991;60:631–652. doi: 10.1146/annurev.bi.60.070191.003215. [DOI] [PubMed] [Google Scholar]
- Gilboa E., Smith C. Gene therapy for infectious diseases: the AIDS model. Trends Genet. 1994 Apr;10(4):139–144. doi: 10.1016/0168-9525(94)90216-x. [DOI] [PubMed] [Google Scholar]
- Green M., Ishino M., Loewenstein P. M. Mutational analysis of HIV-1 Tat minimal domain peptides: identification of trans-dominant mutants that suppress HIV-LTR-driven gene expression. Cell. 1989 Jul 14;58(1):215–223. doi: 10.1016/0092-8674(89)90417-0. [DOI] [PubMed] [Google Scholar]
- Jones K. A., Peterlin B. M. Control of RNA initiation and elongation at the HIV-1 promoter. Annu Rev Biochem. 1994;63:717–743. doi: 10.1146/annurev.bi.63.070194.003441. [DOI] [PubMed] [Google Scholar]
- Luo Y., Madore S. J., Parslow T. G., Cullen B. R., Peterlin B. M. Functional analysis of interactions between Tat and the trans-activation response element of human immunodeficiency virus type 1 in cells. J Virol. 1993 Sep;67(9):5617–5622. doi: 10.1128/jvi.67.9.5617-5622.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Majello B., De Luca P., Hagen G., Suske G., Lania L. Different members of the Sp1 multigene family exert opposite transcriptional regulation of the long terminal repeat of HIV-1. Nucleic Acids Res. 1994 Nov 25;22(23):4914–4921. doi: 10.1093/nar/22.23.4914. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Margolin J. F., Friedman J. R., Meyer W. K., Vissing H., Thiesen H. J., Rauscher F. J., 3rd Krüppel-associated boxes are potent transcriptional repression domains. Proc Natl Acad Sci U S A. 1994 May 10;91(10):4509–4513. doi: 10.1073/pnas.91.10.4509. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Modesti N., Garcia J., Debouck C., Peterlin M., Gaynor R. Trans-dominant Tat mutants with alterations in the basic domain inhibit HIV-1 gene expression. New Biol. 1991 Aug;3(8):759–768. [PubMed] [Google Scholar]
- Pengue G., Calabrò V., Bartoli P. C., Pagliuca A., Lania L. Repression of transcriptional activity at a distance by the evolutionarily conserved KRAB domain present in a subfamily of zinc finger proteins. Nucleic Acids Res. 1994 Aug 11;22(15):2908–2914. doi: 10.1093/nar/22.15.2908. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Pomerantz J. L., Sharp P. A., Pabo C. O. Structure-based design of transcription factors. Science. 1995 Jan 6;267(5194):93–96. doi: 10.1126/science.7809612. [DOI] [PubMed] [Google Scholar]
- Southgate C. D., Green M. R. The HIV-1 Tat protein activates transcription from an upstream DNA-binding site: implications for Tat function. Genes Dev. 1991 Dec;5(12B):2496–2507. doi: 10.1101/gad.5.12b.2496. [DOI] [PubMed] [Google Scholar]
- Tiley L. S., Madore S. J., Malim M. H., Cullen B. R. The VP16 transcription activation domain is functional when targeted to a promoter-proximal RNA sequence. Genes Dev. 1992 Nov;6(11):2077–2087. doi: 10.1101/gad.6.11.2077. [DOI] [PubMed] [Google Scholar]
- Ursini M. V., Lettieri T., Braddock M., Martini G. Enhanced activity of human G6PD promoter transfected in HeLa cells producing high levels of HIV-1 Tat. Virology. 1993 Sep;196(1):338–343. doi: 10.1006/viro.1993.1485. [DOI] [PubMed] [Google Scholar]
- Witzgall R., O'Leary E., Leaf A., Onaldi D., Bonventre J. V. The Krüppel-associated box-A (KRAB-A) domain of zinc finger proteins mediates transcriptional repression. Proc Natl Acad Sci U S A. 1994 May 10;91(10):4514–4518. doi: 10.1073/pnas.91.10.4514. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Yu M., Poeschla E., Wong-Staal F. Progress towards gene therapy for HIV infection. Gene Ther. 1994 Jan;1(1):13–26. [PubMed] [Google Scholar]
