A novel spontaneous mutation of the bovine papillomavirus-1 genome

A cell clone (cl.2) having an atypical transformed morphology was isolated from a murine C127 cell culture experimentally infected with a bovine papillomavirus type 1 (BPV-1) virion preparation. cl.2 cells exhibited minimal transformed characteristics and contained multiple copies of a BPV-1 plasmid with a molecular size slightly less than that of the wild type viral genome. A simple deletion of 277 bp was mapped to the distal portion of the viral 69% transforming fragment where the early gene region merges with the late gene region. None of the recognized early open reading frames were affected by the deletion but sequences including the common early gene mRNA polyadenylation (poly(A] signal and several base pairs of the "distal" enhancer element were deleted. Transfection of C127 cultures with low molecular weight (Hirt) DNA prepared from cl.2 cells led to the appearance of transformed cell foci, and Southern blotting analysis of a cl.2 Hirt DNA-transformed cell line confirmed that the deletion did not destroy the ability to replicate as a high copy plasmid. Removal of the natural early poly(A) signal did not obligate use of the alternative natural viral poly(A) signal located towards the end of the late region. Instead, a new major early mRNA polyadenylation site was mapped close to the unique BamHI recognition sequence at the distal end of the transforming region. Our results underline previous observations that there is a block to the production of stable mRNAs from the late gene region in BPV-1-transformed C127 cells, yet this is not necessarily explained by premature termination of transcription within this region.