Identification of the enhancer binding protein MBF-1 of the sea urchin modulator alpha-H2A histone gene

Biochem Biophys Res Commun. 2002 Jul 12;295(2):519-25. doi: 10.1016/s0006-291x(02)00708-8.

Abstract

The modulator of the sea urchin alpha-H2A histone gene promoter is the only enhancer identified in the alpha-histone gene cluster. Binding of a single factor, denoted MBF-1, has previously detected in nuclear extracts from morula and gastrula embryos. Here, we describe the cloning of MBF-1 by screening a cDNA expression library with a tandem array of modulator binding sites. MBF-1 presents no similarity with other DNA binding proteins and contains nine Krüppel like Zn fingers. In vitro translated proteins and a factor from nuclear extracts interact with the modulator with identical specificity. In addition, MBF-1 expressed in human cells transactivates a reporter gene driven by an array of modulator sites. The DNA binding domain consists of the Zn fingers plus an adjacent basic region, while sequences in the N-terminal region mediates the transactivation function. MBF-1 is expressed in the unfertilized egg and in early and late developmental stages thus confirming that it is not a stage specific enhancer binding factor and that silencing of the alpha-H2A gene after hatching is not due to the lack of the transactivator.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Animals
  • Base Sequence
  • Cloning, Molecular
  • DNA Primers
  • DNA, Complementary
  • DNA-Binding Proteins / chemistry
  • DNA-Binding Proteins / genetics
  • DNA-Binding Proteins / metabolism*
  • Genes, Reporter
  • Histones / genetics*
  • Histones / metabolism
  • Molecular Sequence Data
  • Sea Urchins / genetics*
  • Sequence Homology, Amino Acid
  • Trans-Activators / metabolism

Substances

  • DNA Primers
  • DNA, Complementary
  • DNA-Binding Proteins
  • Histones
  • Trans-Activators

Associated data

  • GENBANK/AJ430050