DNA H form requires a homopurine-homopyrimidine mirror repeat

Regular homopurine-homopyrimidine tracts, (dG-dA)n(dT-dC)n and (dG)n(dC)n, undergo a superhelix-induced, strongly pH-dependent, structural transition into a novel DNA conformation, the H form. We have suggested that the H form can arise in any homopurine-homopyrimidine mirror repeat (H palindrome). We have now tested this prediction using a tailored series of plasmids carrying the inserts AAGGGAGAAXGGGGTATAGGGGYAAGAGGGAA, where X and Y may be either A or G, and subject them to two-dimensional gel electrophoresis. In support of our hypothesis, the inserts exhibited facile transitions into the H form for X = Y = G, or X = Y = A, whereas the transition was much more difficult or impossible for the two non-palindromes (X = A, Y = G or X = G, Y = A). We present evidence that the H form is the structural basis for S1-nuclease hypersensitivity.