Transcriptional regulatory and response mapping of the rat Ha-ras upstream sequence using primary mammary epithelial cells

Carcinogenesis. 1995 Apr;16(4):965-8. doi: 10.1093/carcin/16.4.965.

Abstract

The rat Ha-ras upstream sequence (-2876 to +986 bp relative to the most 5' transcriptional start site) was transcriptionally mapped at a gross level. Ha-ras upstream sequence and 5' unidirectional deletion reporter constructs were transfected via particle bombardment into primary cultures of rat mammary epithelial cells. Analyses of Ha-ras reporter expression show that a fragment extending from -2876 to -2110 bp contains a positive regulatory sequence. The majority of Ha-ras expression is attributed to this sequence, since its deletion results in a 4-fold decrease in expression. The Ha-ras gene was also assessed for autoregulation using similar co-transfection experiments. Wild-type and activated (codon 12 G-->A transition) Ha-ras expression vectors, transcriptionally driven by Ha-ras upstream sequence, were co-transfected with Ha-ras upstream sequence deletion reporter constructs. The activated Ha-ras gene product induced its own transcription 2-fold, targeting the regulatory region between -2119 and -313 bp.

Publication types

  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Animals
  • Base Sequence
  • Cells, Cultured
  • Chromosome Mapping*
  • Epithelium / physiology
  • Female
  • Gene Deletion
  • Genes, Regulator*
  • Genes, ras*
  • Mammary Glands, Animal / physiology*
  • Molecular Sequence Data
  • Mutation
  • Promoter Regions, Genetic
  • Rats
  • Rats, Inbred WF
  • Sequence Analysis, DNA / methods
  • Transcription, Genetic
  • Transfection