Monoclonal antibodies to NF-Y define its function in MHC class II and albumin gene transcription

EMBO J. 1992 Sep;11(9):3315-22.

Abstract

NF-Y is a sequence-specific DNA-binding protein which, as a heterodimer, recognizes CCAAT motifs in a variety of transcriptional promoters. We have generated a panel of monoclonal and affinity-purified polyclonal antibodies directed against various epitopes of NF-Y. These reagents are highly specific for either of the A or B subunits; we have mapped the epitopes recognized by the monoclonal antibodies to the glutamine-rich activation domain of NF-YA. The antibodies inhibit in vitro transcription from the promoters of the albumin gene and of Ea, a class II gene of the major histocompatibility complex. These data definitively demonstrate the role of NF-Y in regulating the transcription of two tissue-specific genes whose expression patterns do not overlap. Interestingly, the antibodies cannot inhibit a formed pre-initiation complex, but do block reinitiation of subsequent rounds of transcription from the same templates.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Albumins / biosynthesis
  • Albumins / genetics*
  • Animals
  • Antibodies, Monoclonal / immunology
  • Antibody Affinity
  • Antibody Specificity
  • Base Sequence
  • CCAAT-Enhancer-Binding Proteins
  • Cloning, Molecular
  • DNA-Binding Proteins / immunology
  • DNA-Binding Proteins / physiology*
  • Epitopes
  • Gene Expression Regulation*
  • Genes, MHC Class II*
  • Mice
  • Mice, Inbred BALB C
  • Models, Genetic
  • Molecular Sequence Data
  • Transcription Factors / immunology
  • Transcription Factors / physiology*
  • Transcription, Genetic

Substances

  • Albumins
  • Antibodies, Monoclonal
  • CCAAT-Enhancer-Binding Proteins
  • DNA-Binding Proteins
  • Epitopes
  • Transcription Factors