Transfer of murine intracisternal A particle phenotype in chloramphenicol-resistant cytoplasts

Abstract

Murine intracisternal A particles have a number of properties which are common to known RNA tumor viruses, but horizontal transmission has not been previously demonstrated. The apparent absence of infectivity may be related to the failure of these particles to be released from cisternae of endo-plasmic reticulum. Previous biological studies using isolated, purified A particles have been compromised by the fact that the isolation procedure requires small amounts of nonionic detergent. Using some techniques of somatic cell hybridization, we have assessed the capacity for A particle genome transfer from positive to negative cells. Since it has been previously shown that some chloramphenicol-resistant cell lines can transfer this resistance in the cytoplasm, we have used this characteristic as a marker for cytoplasmic fragments. Mouse cells containing A particles were mutagenized, and clones resistant to chloramphenicol were selected; by enucleating these cells and fusing the resultant cytoplasts to each of two recipient mouse cell lines negative for A particles, it is possible to identify clones of cells known to be the product of a fusion event between a cytoplast and a whole cell (cybrids). Under these conditions, intracisternal A particles appear in the cybrid clones as a phenotypic trait that has not been segregated over at least 60-80 cell generations.