Purification of a nuclear trans-acting factor involved in the regulated transcription of a human immunoglobulin heavy chain gene

Cell. 1988 Jun 3;53(5):723-30. doi: 10.1016/0092-8674(88)90090-6.

Abstract

The expression of the rearranged human immunoglobulin gamma 1 heavy chain gene (HIG1) was shown to be induced through its enhancer by the positive regulatory trans-acting factor(s) that was contained only in cells of B lineage. The trans-acting factors were purified from mouse myeloma NS1 cells, and HIG1-inducing activity was found mainly in fractions of molecular weight 53-127 kd and in a fraction eluted from a heparin-Sepharose column with 0.5 M KCI. This semipurified fraction contained proteins binding to the conserved octamer sequence, ATGCAAAT, in the promoter region, as well as to sequences in the enhancer region. The 0.5 M KCI eluates from a heparin-Sepharose column were applied to a DNA affinity column of synthetic oligonucleotides of the octamer sequence and the sequence TATTTTAGGAAGCAAA in the HpaII-BgIII region of the HIG1 gene enhancer. The protein eluted from the enhancer sequence-specific DNA affinity column showed a strong inducing activity for the HIG1 gene, and the molecular weight of a predominant protein was 96 kd.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Base Sequence
  • Cell Line
  • Cell Nucleus / metabolism*
  • DNA Restriction Enzymes
  • Enhancer Elements, Genetic*
  • Gene Expression Regulation*
  • Gene Products, tat
  • Genes, Immunoglobulin*
  • Humans
  • Immunoglobulin Heavy Chains / genetics*
  • Mice
  • Molecular Sequence Data
  • Molecular Weight
  • Transcription Factors / isolation & purification*
  • Transcription Factors / physiology
  • Transcription, Genetic*

Substances

  • Gene Products, tat
  • Immunoglobulin Heavy Chains
  • Transcription Factors
  • DNA Restriction Enzymes