Ternary complex formation between MADS-box transcription factors and the histone fold protein NF-YB

J Biol Chem. 2002 Jul 19;277(29):26429-35. doi: 10.1074/jbc.M202546200. Epub 2002 Apr 23.

Abstract

MADS-box proteins are transcription factors present in different eukaryotic kingdoms. In contrast to plants, for mammalian and yeast MADS-box proteins ternary complex formation with unrelated transcription factors was reported. We show here the first identification of such ternary interaction in plants. A rice seed-specific NF-YB was identified as partner of OsMADS18 by two-hybrid screening. NF-YB contains a histone fold motif, HFM,(1) and is part of the trimeric CCAAT-binding NF-Y complex. OsMADS18, alone or in combination with a natural partner, interacts with OsNF-YB1 through the MADS and I regions. The mouse NF-YB also associates with OsMADS18 in vivo and in vitro as a NF-YB-NF-YC dimer. Other rice MADS-box proteins do not interact in these assays, indicating specificity for the interaction. OsNF-YB1 is capable of heterodimerizing with NF-YC, but not trimerizing with NF-YA, thus precluding CCAAT binding. Mutation of the variant Asp at position 99 of the HFM alpha2-helix into a conserved serine recovers the capacity to interact with NF-YA, but not with DNA. This is the first indication that members of the NF-YB family work through mechanisms independent of the CCAAT box.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Animals
  • Arabidopsis
  • CCAAT-Binding Factor / metabolism*
  • DNA-Binding Proteins / metabolism*
  • MADS Domain Proteins / metabolism*
  • Mice
  • Molecular Sequence Data
  • Plant Proteins / metabolism*
  • Protein Binding
  • Protein Conformation
  • Protein Structure, Secondary
  • Rabbits
  • Reverse Transcriptase Polymerase Chain Reaction
  • Sequence Alignment
  • Transcription Factors*
  • Two-Hybrid System Techniques

Substances

  • CCAAT-Binding Factor
  • DNA-Binding Proteins
  • MADS Domain Proteins
  • Nfya protein, mouse
  • Nfyb protein, mouse
  • Plant Proteins
  • Transcription Factors