Pseudouridine excretion and transfer RNA primers for reverse transcriptase in tumors of retroviral origin

Abstract

To evaluate the relationship between pseudouridine increase in biological fluids and retroviral cell transformation, we have studied the effect of retrovirus infection and/or transformation on the rate of pseudouridine excretion by chick embryo fibroblasts. The results show that: pseudouridine excretion by chick embryo fibroblasts transformed by Rous sarcoma virus is several times higher than that of normal cells; this increased excretion precedes by many hours the appearance of the morphological signs of transformation and it is always present when neosynthesized infectious viral particles are released into the culture medium; and pseudouridine excretion was also increased in cells infected by a mutant of Rous sarcoma virus (RAV-1) which, lacking the src gene, does not transform the cells but replicates normally. To investigate if pseudouridine overproduction is related to an altered turnover rate of specific transfer RNA (tRNA) species which functions as primer of retrovirus reverse transcriptase, the concentration of non-acylated proline-accepting tRNA and non-acylated tryptophan-accepting tRNA, primers of reverse transcriptase of murine leukemia virus and of Rous sarcoma virus, respectively, has been measured, the former in normal and transformed AKR thymus and the latter in normal fibroblasts and in fibroblasts infected by Rous sarcoma virus or by its nontransforming mutant. The results show that in both systems a significant increase of the primer tRNA species occurs in the infected or transformed cells.