Enhancer functions in U3 of Akv virus: a role for cooperativity of a tandem repeat unit and its flanking DNA sequences

Abstract

cis-Acting transcriptional control elements in the U3 region of the murine retrovirus Akv were analyzed in mouse NIH 3T3 fibroblast cells by using a transient expression vector system based upon a complete long terminal repeat with linked flanking sequences. Deletion analysis pointed to the essential role of sequences within the 99-base-pair direct repeats, and a fragment encompassing the two repeats was found to possess orientation-independent enhancer activity when positioned either upstream or downstream of the transcriptional unit. Removal of one copy of the 99-base-pair repeat led to a reduction in activity of about 2.5-fold when located in an intact U3 environment but to reductions of up to 2 orders of magnitude when placed in other sequence contexts. Our studies of enhancer functions in the presence of one versus two copies of the tandem repeat point to duplicate functions of repeat sequences and sequences flanking the repeat region and emphasize the complex overall organization of this U3 region.