Sequence analysis of the putative viral enhancer in tissues from 33 cats with various feline leukemia virus-related...

Diseases resulting from infection by feline leukemia virus (FeLV) and several other retroviruses relate in part, to non-coding regulatory sequences within the viral long terminal repeat (LTR). Both enhancer repeats and mutations within the LTR have been implicated in FeLV related disease. In order to investigate the relationship between nucleotide sequence of the FeLV LTR and disease, tissues from 33 cats with different types of degenerative and proliferative FeLV-related disease were studied. An FeLV LTR region containing the putative transcriptional enhancer unit was amplified by polymerase chain reaction (PCR) from FeLV-infected tissues. Phylogenetic analysis of FeLV 3'unique (U3) sequences revealed only one meaningful grouping which contained 4 of the 5 antigen-negative lymphosarcomas (LSAs). No sequence duplications were found in any of the 33 FeLV U3 regions. Point mutations relative to the corresponding region of FeLV-A/Glasgow, were identified at 102 positions; 68 of these were accounted for by mutations at 5 locations. Only 1 point mutation was found within the leukemia virus b-simian virus 40-like core (LVb-CORE) site. However, the nuclear factor 1 (NF1) site contained 11 mutations, and the FeLV-specific (FLV-1) site contained 26 mutations. Most of the remaining mutations were upstream of the LVB site between glucocorticoid response element (GRE) and FLV-1. The 10 LSAs, particularly the 5 antigen-negative LSAs, deviated least from the corresponding sequence for FeLV-A/Glasgow. Conclusions were that the spectrum of neoplastic and non-neoplastic FeLV-related diseases investigated in this study, developed in the presence of FeLVs containing the single enhancer unit. The significance of the point mutations is unknown, however, those occurring with high frequency and within nuclear protein binding should be first to be investigated in functional studies.