Structural alteration from non-B to B-form could reflect DNase I hypersensitivity.
Preferential cleavage of active genes by DNase I has been correlated with a structurally altered conformation of DNA at the hypersensitive site in chromatin. To have a better understanding of the structural requirements for gene activation as probed by DNase I action, digestability by DNase I of synthetic polynucleotides having the ability to adopt B and non-B conformation (like Z-form) was studied which indicated a marked higher digestability of the B-form of DNA. Left handed Z form present within a natural sequence in supercoiled plasmid also showed marked resistance towards DNase I digestion. We show that alternating purine-pyrimidine sequences adopting Z-conformation exhibit DNAse I foot printing even in a protein free system. The logical deductions from the results indicate that 1) altered structure like Z-DNA is not a favourable substrate for DNase I, 2) both the ends of the alternating purine-pyrimidine insert showed hypersensitivity, 3) B-form with a minor groove of 12-13 A is a more favourable substrate for DNase I than an altered structure, 4) any structure of DNA deviating largely from B form with a capacity to flip over to the B-form are potential targets for the DNase I enzymic probes in naked DNA.
Ramesh
N
N
Molecular Biophysics Unit, Indian Institute of Science, Bangalore.
Brahmachari
S K
SK
D004247:DNA / Q000378:metabolism*; D003850:Deoxyribonuclease I / Q000502:physiology; D008968:Molecular Conformation; D009710:Nucleotide Mapping; D010957:Plasmids
D004247:DNA; D003850:Deoxyribonuclease I