Detection of the myristylated gag-raf transforming protein with raf-specific antipeptide sera

Virology. 1985 Oct 15;146(1):78-89. doi: 10.1016/0042-6822(85)90054-6.

Abstract

The post-translational modifications of the gag-raf fusion proteins of the 3611 murine sarcoma virus (MSV) have been examined by inhibiting glycosylation with tunicamycin and by in vivo labeling with [3H]myristic acid. The results show that P75gag-raf is myristylated but not glycosylated and that P90gag-raf is glycosylated but not myristylated (and is now termed gP90gag-raf). gP90gag-raf expression appeared to become lost during passage of the transformed cells, and consequently does not appear to be necessary for the maintenance of transformation. raf-specific sera for detecting gag-raf fusion proteins have been obtained from synthetic peptides made from different regions of the predicted v-raf sequence. Immunoprecipitation of P75gag-raf with raf-specific sera directly confirmed the deduced v-raf sequence. The fact that P75gag-raf is both myristylated and precipitated by antiserum to a predicted carboxyl-terminal peptide of the v-raf gene established that the mature protein represents the entire coding region. The gP90gag-raf thus appears to be a glycosylated form of P75gag-raf specified by the gag sequences of the fusion protein, in analogy with Pr65gag and gPr80gag of murine leukemia viruses. Antiserum to the carboxyl-terminal P75gag-raf peptide was the most efficient in immunoprecipitation, and will be useful for detecting the product of the c-raf gene.

Publication types

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

MeSH terms

  • Animals
  • Antigens, Polyomavirus Transforming
  • Antigens, Viral, Tumor / analysis*
  • Antigens, Viral, Tumor / immunology
  • Antigens, Viral, Tumor / metabolism
  • Cell Line
  • Cell Transformation, Neoplastic
  • Cell Transformation, Viral
  • Genes, Viral
  • Immune Sera
  • Mice
  • Molecular Weight
  • Myristic Acid
  • Myristic Acids / metabolism
  • Oncogenes
  • Protein Processing, Post-Translational*
  • Sarcoma Viruses, Murine / analysis*
  • Sarcoma Viruses, Murine / genetics
  • Sarcoma Viruses, Murine / metabolism
  • Viral Envelope Proteins / analysis*
  • Viral Envelope Proteins / metabolism
  • Viral Fusion Proteins
  • Viral Proteins / analysis*
  • Viral Proteins / immunology
  • Viral Proteins / metabolism

Substances

  • Antigens, Polyomavirus Transforming
  • Antigens, Viral, Tumor
  • Immune Sera
  • Myristic Acids
  • Viral Envelope Proteins
  • Viral Fusion Proteins
  • Viral Proteins
  • Myristic Acid