Analysis of the human HLA-DRA gene upstream region: Evidence for a stem-loop array directed by nuclear factors

Carlo Mischiati, Giordana Feriotto, Nicoletta Bianchi, Cristina Rutigliano, Patrizio Giacomini, Roberto Gambari

Research output: Contribution to journalArticle

4 Citations (Scopus)

Abstract

Sequence analysis of the far-upstream region of the human HLA-DRA gene has revealed the presence of Y' and X' boxes, highly homologous to the well characterized Y and X boxes present within the proximal-promoter region. Comparison of Y, Y', X, and X' box sequences present within different class II MHC genes of different species demonstrates that these boxes are conserved during evolution, suggesting an important role in regulation of gene expression. The far-upstream region and the proximal promoter region of the class II MHC genes could be organized in secondary structures, as suggested for the EA gene, the murine counterpart of the human HLA-DRA gene. The essential feature of this model is a dimerization of the proteins binding to X and X' and/or Y and Y' boxes resulting in a loop-out of the intervening DNA and a rapprochement of the far-upstream and proximal-promoter regions, and consequently of any proteins binding to them. We set up an in vitro approach in order to determine whether proteins bound to sequences present within far-upstream and proximal-promoter regions of the human HLA-DRA gene could direct a secondary structure assembly of regulative regions. Moreover, by gel retardation and DNase I footprinting assays, we demonstrate that similar proteins bind to Y and Y' boxes and, among these proteins, NF-Y was unambiguously identified by antibody-super shift experiments. Taken together, the data presented in this paper provide evidence supporting the hypothesis that a stem-loop array of the 5'-upstream region of the human HLA-DRA gene could be directed by nuclear factors. In this manner, additional nuclear factors bound to the far region could be driven in close proximity of the transcription initiation site.

Original languageEnglish
Pages (from-to)219-228
Number of pages10
JournalBiochimie
Volume81
Issue number3
DOIs
Publication statusPublished - Mar 1999

Fingerprint

HLA-DR alpha-Chains
Genetic Promoter Regions
Genes
MHC Class II Genes
Protein Binding
CCAAT-Binding Factor
Transcription Initiation Site
Deoxyribonuclease I
Gene Expression Regulation
Dimerization
Sequence Analysis
Proteins
Gels
Gene expression
Assays
Antibodies
DNA

Keywords

  • HLA-DRA gene
  • NF-Y
  • Promoter

ASJC Scopus subject areas

  • Biochemistry

Cite this

Analysis of the human HLA-DRA gene upstream region : Evidence for a stem-loop array directed by nuclear factors. / Mischiati, Carlo; Feriotto, Giordana; Bianchi, Nicoletta; Rutigliano, Cristina; Giacomini, Patrizio; Gambari, Roberto.

In: Biochimie, Vol. 81, No. 3, 03.1999, p. 219-228.

Research output: Contribution to journalArticle

Mischiati, Carlo ; Feriotto, Giordana ; Bianchi, Nicoletta ; Rutigliano, Cristina ; Giacomini, Patrizio ; Gambari, Roberto. / Analysis of the human HLA-DRA gene upstream region : Evidence for a stem-loop array directed by nuclear factors. In: Biochimie. 1999 ; Vol. 81, No. 3. pp. 219-228.
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