Identification of ETS domain proteins in murine T lymphocytes that interact with the Moloney murine leukemia virus enhancer
- PMID: 7935472
- PMCID: PMC359293
- DOI: 10.1128/mcb.14.11.7569-7580.1994
Identification of ETS domain proteins in murine T lymphocytes that interact with the Moloney murine leukemia virus enhancer
Abstract
The enhancer of Moloney murine leukemia virus (Mo-MuLV) contains an array of transcriptional control elements that direct viral gene expression in diverse cell types. The murine transcription factor Ets-1 was shown to bind to the LVb and LVc elements of the enhancer by DNase I protection and methylation interference assays. Enhancers containing disrupted Ets-1 binding sites were tested in transient expression assays in the murine T-cell line EL4.E1; alterations in the LVb element affected constitutive enhancer activity, while mutation of either the LVb or LVc element disrupted phorbol ester-induced enhancer activity. Members of the ets gene family of proteins display similar DNA-binding properties; therefore, we speculated that ets proteins other than Ets-1 also might bind these elements. Crude nuclear extracts of EL4.E1 cells were assayed to identify the protein(s) that potentially functions at the LVb element. The predominant binding activity was not Ets-1 but rather two independent DNA-protein complexes that comigrated in mobility shift assays. UV cross-linking and denaturing gel electrophoresis sized the two DNA-binding species, which we denoted p55 and p100. Immunoprecipitation combined with UV cross-linking identified p55 as the alpha subunit of GA-binding protein. The DNA-binding properties of p100 and several ets proteins were compared. Similarities suggested that p100 is also an ETS domain protein, possibly Elf-1. This strategy could be used to identify other ETS domain proteins in crude nuclear extracts. These findings suggest multiple ETS domain proteins could regulate gene expression of Mo-MuLV.
Similar articles
-
Transactivation of the Moloney murine leukemia virus and T-cell receptor beta-chain enhancers by cbf and ets requires intact binding sites for both proteins.J Virol. 1995 Aug;69(8):4941-9. doi: 10.1128/JVI.69.8.4941-4949.1995. J Virol. 1995. PMID: 7609063 Free PMC article.
-
cis-acting sequences required for inducible interleukin-2 enhancer function bind a novel Ets-related protein, Elf-1.Mol Cell Biol. 1992 Mar;12(3):1043-53. doi: 10.1128/mcb.12.3.1043-1053.1992. Mol Cell Biol. 1992. PMID: 1545787 Free PMC article.
-
In vivo footprinting of the enhancer sequences in the upstream long terminal repeat of Moloney murine leukemia virus: differential binding of nuclear factors in different cell types.J Virol. 1998 Nov;72(11):8961-70. doi: 10.1128/JVI.72.11.8961-8970.1998. J Virol. 1998. PMID: 9765441 Free PMC article.
-
Genomic and biochemical insights into the specificity of ETS transcription factors.Annu Rev Biochem. 2011;80:437-71. doi: 10.1146/annurev.biochem.79.081507.103945. Annu Rev Biochem. 2011. PMID: 21548782 Free PMC article. Review.
-
Enhancer dysfunction in leukemia.Blood. 2018 Apr 19;131(16):1795-1804. doi: 10.1182/blood-2017-11-737379. Epub 2018 Feb 9. Blood. 2018. PMID: 29439951 Free PMC article. Review.
Cited by
-
Expression profiles frame the promoter specificity dilemma of the ETS family of transcription factors.Nucleic Acids Res. 2004 Oct 21;32(18):5693-702. doi: 10.1093/nar/gkh906. Print 2004. Nucleic Acids Res. 2004. PMID: 15498926 Free PMC article.
-
Methylation of an ETS site in the intron enhancer of the keratin 18 gene participates in tissue-specific repression.Mol Cell Biol. 1997 Sep;17(9):4885-94. doi: 10.1128/MCB.17.9.4885. Mol Cell Biol. 1997. PMID: 9271368 Free PMC article.
-
Members of the GATA family of transcription factors bind to the U3 region of Cas-Br-E and graffi retroviruses and transactivate their expression.J Virol. 1998 Jul;72(7):5579-88. doi: 10.1128/JVI.72.7.5579-5588.1998. J Virol. 1998. PMID: 9621016 Free PMC article.
-
RUNX1 and RUNX1-ETO: roles in hematopoiesis and leukemogenesis.Front Biosci (Landmark Ed). 2012 Jan 1;17(3):1120-39. doi: 10.2741/3977. Front Biosci (Landmark Ed). 2012. PMID: 22201794 Free PMC article. Review.
-
A potential role for Elf-1 in terminal transferase gene regulation.Mol Cell Biol. 1996 Nov;16(11):6121-31. doi: 10.1128/MCB.16.11.6121. Mol Cell Biol. 1996. PMID: 8887642 Free PMC article.
References
Publication types
MeSH terms
Substances
Grants and funding
LinkOut - more resources
Full Text Sources
Molecular Biology Databases
Miscellaneous