Variants of JC virus (JCV) strain GS were isolated directly from the central nervous system (variant GS/B) and the kidney (variant GS/K) of a patient with progressive multifocal leukoencephalopathy and were cloned and sequenced. The genomes of the isolates were shown to be nearly identical in the nucleotide sequences of their protein-coding regions, suggesting that both had originated from a single infecting JCV genome. In contrast, the arrangement of the putative elements of transcriptional control revealed considerable differences. The tandemly repeated elements found twice within the enhancer region of JCV GS/B variant were not present in the GS/K variant. The missing elements were replaced by DNA segments containing simian virus 40 and adenovirus E1A core enhancer elements. These differences in the organ-specific GS variants suggest that rearrangements within elements of transcriptional control might be involved in altering the virus-cell interaction in the course of a JCV infection.