Inhibitory effect of the transcription factor encoded by the mi mutant allele in cultured mast cells of mice

Blood. 1999 Feb 15;93(4):1189-96.

Abstract

The mi locus of mice encodes a transcription factor of the basic-helix-loop-helix-leucine zipper protein family (MITF). The MITF encoded by the mutant mi allele (mi-MITF) deletes 1 of 4 consecutive arginines in the basic domain. The mice of mi/mi genotype express mi-MITF, whereas the mice of tg/tg genotype have a transgene at the 5' flanking region of the mi gene and do not express any MITF. To investigate the function of mi-MITF in cultured mast cells (CMCs), we took two approaches. First, mRNA obtained from mi/mi CMCs or tg/tg CMCs was subtracted from complementary (c) DNA library of normal (+/+) CMCs, and the (+/+-mi/mi) and (+/+-tg/tg) subtraction libraries were obtained. When the number of clones that hybridized more efficiently with +/+ CMC cDNA probe than with mi/mi or tg/tg CMC cDNA probe was compared using Southern analysis, the number was larger in the (+/+-mi/mi) library than in the (+/+-tg/tg) library. Second, we compared mRNA expression of six genes between mi/mi and tg/tg CMCs by Northern analysis. The transcription of three genes encoding mouse mast cell proteases was impaired in both mi/mi and tg/tg CMCs. On the other hand, the transcription of three genes encoding c-kit receptor, tryptophan hydroxylase, and granzyme B was markedly reduced in mi/mi CMCs, but the reduction was significantly smaller in tg/tg CMCs. These results indicated the inhibitory effect of mi-MITF on the transactivation of particular genes in CMCs.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Cells, Cultured
  • DNA-Binding Proteins / genetics*
  • Gene Expression Regulation*
  • Helix-Loop-Helix Motifs
  • Leucine Zippers
  • Mast Cells / physiology*
  • Mice
  • Mice, Inbred C57BL
  • Microphthalmia-Associated Transcription Factor
  • Mutation
  • Transcription Factors / genetics*

Substances

  • DNA-Binding Proteins
  • Microphthalmia-Associated Transcription Factor
  • Mitf protein, mouse
  • Transcription Factors