Subtraction cloning of H-rev107, a gene specifically expressed in H-ras resistant fibroblasts

Oncogene. 1994 Feb;9(2):479-90.

Abstract

We have isolated by subtractive hybridization a novel gene, called H-rev107, which is specifically expressed in a phenotypic revertant of H-ras transformed 208F rat fibroblasts. Apart from oncogene revertants, strong expression of H-rev107 was found in REF52 and EK-3 cells, two fibroblast lines resistant to transformation by activated H-ras oncogenes. In contrast, transformation-sensitive fibroblasts like 208F or NIH3T3 cells expressed only very little H-rev107 RNA. In H-ras or v-src transformed fibroblasts, H-rev107 RNA was undetectable. Introduction of the adenovirus E1A nuclear oncogene into ras-resistant REF52 cells abolished their transformation resistance and repressed the H-rev107 gene. H-rev107 encodes a protein with a molecular weight of 18 kDa without any structural similarity to known proteins. p18H-rev107 exists in two forms which can be distinguished by their electrophoretic mobility; one is localized predominantly in cell membranes, the other in the cytoplasm. In confluent contact-inhibited 208F cells, p18H-rev107 accumulated in cell membranes, while growth arrest induced by serum starvation did not induce H-rev107. In REF52, cell density had no influence on the expression or localization of p18H-rev107. Repression of the H-rev107 gene may be closely associated with the loss of density-dependent growth inhibition and with the expression of the neoplastic phenotype.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adenovirus E1A Proteins / physiology
  • Amino Acid Sequence
  • Animals
  • Base Sequence
  • Blotting, Northern
  • Blotting, Western
  • Cell Line
  • Cell Membrane / chemistry
  • Cell Transformation, Neoplastic / genetics*
  • Cell Transformation, Neoplastic / pathology
  • Cloning, Molecular
  • Cytoplasm / chemistry
  • DNA / analysis
  • DNA / genetics
  • Down-Regulation / physiology
  • Fibroblasts / chemistry
  • Fibroblasts / metabolism
  • Fibroblasts / pathology*
  • Genes, ras / genetics
  • Genes, ras / physiology*
  • Molecular Sequence Data
  • Molecular Weight
  • Phenotype
  • Phospholipases A2, Calcium-Independent
  • Proteins / analysis
  • Proteins / genetics*
  • Proteins / metabolism
  • RNA, Messenger / analysis
  • RNA, Messenger / genetics
  • Rats
  • Tumor Suppressor Proteins

Substances

  • Adenovirus E1A Proteins
  • Proteins
  • RNA, Messenger
  • Tumor Suppressor Proteins
  • DNA
  • Phospholipases A2, Calcium-Independent
  • Plaat3 protein, rat