Interactions of the nuclear matrix-associated steroid receptor binding factor with its DNA binding element in the c-myc gene promoter

Thomas J. Barrett, Nicole P. Sandhu, Andy J. Tomlinson, Linda M. Benson, Malayannan Subramaniam, Stephen Naylor, Thomas C. Spelsberg

Research output: Contribution to journalArticle

9 Citations (Scopus)

Abstract

Steroid receptor binding factor (RBF) was originally isolated from avian oviduct nuclear matrix. When bound to avian genomic DNA, RBF generates saturable high-affinity binding sites for the avian progesterone receptor (PR). Recent studies have shown that RBF binds to a 54 bp element in the 5'- flanking region of the progesterone-regulated avian c-myc gene, and nuclear matrix-like attachment sites flank the RBF element [Lauber et al. (1997) J. Biol. Chem. 272, 24657-24665]. In this paper, electrophoretic mobility shift assays (EMSAs) and S1 nuclease treatment are used to demonstrate that the RBF-maltose binding protei (MBP) fusion protein binds to single-stranded DNA of its element. Only the N-terminal domain of RBF binds the RBF DNA element as demonstrated by southwestern blot analyses, and by competition EMSAs between RBF-MBP and the N-terminal domain. Mass spectrometric analysis of the C-terminal domain of RBF demonstrates its potential to form noncovalent protein-protein interactions via a potential leucine-isoleucine zipperlike structure, suggesting a homo- and/or possible heterodimer structure in solution. These data support that the nuclear matrix binding site (acceptor site) for PR in the c-myc gene promoter is composed of RBF dimers bound to a specific single-stranded DNA element. The dimers of RBF are generated by C- terminal leucine zipper and the DNA binding occurs at the N-terminal parallel β-sheet DNA binding motif. This complex is flanked by nuclear matrix attachment sites.

Original languageEnglish (US)
Pages (from-to)753-762
Number of pages10
JournalBiochemistry
Volume39
Issue number4
DOIs
StatePublished - Feb 1 2000

Fingerprint

Nuclear Matrix
myc Genes
Steroid Receptors
Electrophoretic mobility
Maltose
Genes
Single-Stranded DNA
Progesterone Receptors
Dimers
Proteus
Assays
DNA
Electrophoretic Mobility Shift Assay
Binding Sites
Leucine Zippers
Proteins
Southwestern Blotting
Isoleucine
5' Flanking Region
Leucine

ASJC Scopus subject areas

  • Biochemistry

Cite this

Barrett, T. J., Sandhu, N. P., Tomlinson, A. J., Benson, L. M., Subramaniam, M., Naylor, S., & Spelsberg, T. C. (2000). Interactions of the nuclear matrix-associated steroid receptor binding factor with its DNA binding element in the c-myc gene promoter. Biochemistry, 39(4), 753-762. https://doi.org/10.1021/bi991809v

Interactions of the nuclear matrix-associated steroid receptor binding factor with its DNA binding element in the c-myc gene promoter. / Barrett, Thomas J.; Sandhu, Nicole P.; Tomlinson, Andy J.; Benson, Linda M.; Subramaniam, Malayannan; Naylor, Stephen; Spelsberg, Thomas C.

In: Biochemistry, Vol. 39, No. 4, 01.02.2000, p. 753-762.

Research output: Contribution to journalArticle

Barrett, TJ, Sandhu, NP, Tomlinson, AJ, Benson, LM, Subramaniam, M, Naylor, S & Spelsberg, TC 2000, 'Interactions of the nuclear matrix-associated steroid receptor binding factor with its DNA binding element in the c-myc gene promoter', Biochemistry, vol. 39, no. 4, pp. 753-762. https://doi.org/10.1021/bi991809v
Barrett, Thomas J. ; Sandhu, Nicole P. ; Tomlinson, Andy J. ; Benson, Linda M. ; Subramaniam, Malayannan ; Naylor, Stephen ; Spelsberg, Thomas C. / Interactions of the nuclear matrix-associated steroid receptor binding factor with its DNA binding element in the c-myc gene promoter. In: Biochemistry. 2000 ; Vol. 39, No. 4. pp. 753-762.
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