Cloning of a negative transcription factor that binds to the upstream conserved region of moloney murine leukemia virus

James R. Flanagan, Kevin G. Becker, David L. Ennist, Shannon L. Gleason, Paul H. Driggers, Ben Zion Levi, Ettore Appella, Keiko Ozato

Research output: Contribution to journalArticle

Abstract

The long terminal repeat of Moloney murine leukemia virus (MuLV) contains the upstream conserved region (UCR). The UCR core sequence, CGCCATTTT, binds a ubiquitous nuclear factor and mediates negative regulation of MuLV promoter activity. We have isolated murine cDNA clones encoding a protein, referred to as UCRBP, that binds specifically to the UCR core sequence. Gel mobility shift assays demonstrate that the UCRBP fusion protein expressed in bacteria binds the UCR core with specificity identical to that of the UCR-binding factor in the nucleus of murine and human cells. Analysis of full-length UCRBP cDNA reveals that it has a putative zinc finger domain composed of four C2H2 zinc fingers of the GLI subgroup and an N-terminal region containing alternating charges, including a stretch of 12 histidine residues. The 2.4-kb UCRBP message is expressed in all cell lines examined (teratocarcinoma, B- and T-cell, macrophage, fibroblast, and myocyte), consistent with the ubiquitous expression of the UCR-binding factor. Transient transfection of an expressible UCRBP cDNA into fibroblasts results in down-regulation of MuLV promoter activity, in agreement with previous functional analysis of the UCR. Recently three groups have independently isolated human and mouse UCRBP. These studies show that UCRBP binds to various target motifs that are distinct from the UCR motif: the adeno-associated virus PS promoter and elements in the immunoglobulin light- and heavy-chain genes, as well as elements in ribosomal protein genes. These results indicate that UCRBP has unusually diverse DNA-binding specificity and as such is likely to regulate expression of many different genes.

Original languageEnglish (US)
Pages (from-to)38-44
Number of pages7
JournalMolecular and Cellular Biology
Volume12
Issue number1
StatePublished - Jan 1992
Externally publishedYes

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Moloney murine leukemia virus
Organism Cloning
Murine Leukemia Viruses
Transcription Factors
Complementary DNA
Immunoglobulin Light Chain Genes
Fibroblasts
Teratocarcinoma
Immunoglobulin Heavy Chain Genes
Dependovirus
Terminal Repeat Sequences
Ribosomal Proteins
Zinc Fingers
Electrophoretic Mobility Shift Assay
Histidine
Muscle Cells
Genes
Transfection
Proteins
Down-Regulation

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics
  • Cell Biology

Cite this

Flanagan, J. R., Becker, K. G., Ennist, D. L., Gleason, S. L., Driggers, P. H., Levi, B. Z., ... Ozato, K. (1992). Cloning of a negative transcription factor that binds to the upstream conserved region of moloney murine leukemia virus. Molecular and Cellular Biology, 12(1), 38-44.

Cloning of a negative transcription factor that binds to the upstream conserved region of moloney murine leukemia virus. / Flanagan, James R.; Becker, Kevin G.; Ennist, David L.; Gleason, Shannon L.; Driggers, Paul H.; Levi, Ben Zion; Appella, Ettore; Ozato, Keiko.

In: Molecular and Cellular Biology, Vol. 12, No. 1, 01.1992, p. 38-44.

Research output: Contribution to journalArticle

Flanagan, JR, Becker, KG, Ennist, DL, Gleason, SL, Driggers, PH, Levi, BZ, Appella, E & Ozato, K 1992, 'Cloning of a negative transcription factor that binds to the upstream conserved region of moloney murine leukemia virus', Molecular and Cellular Biology, vol. 12, no. 1, pp. 38-44.
Flanagan JR, Becker KG, Ennist DL, Gleason SL, Driggers PH, Levi BZ et al. Cloning of a negative transcription factor that binds to the upstream conserved region of moloney murine leukemia virus. Molecular and Cellular Biology. 1992 Jan;12(1):38-44.
Flanagan, James R. ; Becker, Kevin G. ; Ennist, David L. ; Gleason, Shannon L. ; Driggers, Paul H. ; Levi, Ben Zion ; Appella, Ettore ; Ozato, Keiko. / Cloning of a negative transcription factor that binds to the upstream conserved region of moloney murine leukemia virus. In: Molecular and Cellular Biology. 1992 ; Vol. 12, No. 1. pp. 38-44.
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