Photofootprint of nucleosome core DNA in intact chromatin having different structural states.
Recently, we reported that the distribution of ultraviolet light (u.v.) induced pyrimidine dimers in nucleosome core DNA has a striking 10.3(+/- 0.1) base periodicity and the regions of enhanced quantum yield map to positions where DNA strands are farthest from the core histone surface. Improvement of the mapping procedure has allowed us to analyze this distribution in more detail, and compare the distribution pattern for nucleosome cores from intact chromatin having different higher-order structures (from the 10 nm filament to the 30 nm fiber). At all levels of chromatin compaction, we observed the following. (1) The average periodicity in pyrimidine dimer yield is 10.3 bases. (2) The peak-to-peak spacing in this distribution is significantly different from 10.3 bases in the region covering three helix turns immediately 5' of the dyad axis. (3) There is a suppression of photoproduct formation in the region of the dyad axis, especially at position 84 from the 5' end. (4) The approximately 10 base ensembles have alternating peak intensities throughout core DNA. Furthermore, peak deconvolution analysis of the pyrimidine dimer pattern yielded a striking similarity in photoproduct yield for the different levels of chromatin compaction. Irradiation of isolated core DNA yields a much more random distribution of photoproducts, although a weak modulation pattern is observed (indicating that there is a non-random alignment of adjacent pyrimidines in our core DNA preparations). This pattern includes a depression in photoproduct yield near position 95, suggesting that the sequence in this region plays a role in nucleosome positioning. These results show that the u.v. photofootprint is a sensitive, diagnostic probe of core histone-DNA interactions in intact chromatin, and these interactions are not significantly altered by changes in the structural state of the chromatin fiber.
Gale
J M
JM
Biochemistry/Biophysics Program, Washington State University, Pullman 99164-4660.
Smerdon
M J
MJ
D000818:Animals; D002417:Cattle; D002843:Chromatin / Q000235:genetics* / Q000378:metabolism; D004247:DNA / Q000235:genetics*; D003720:Densitometry; D006657:Histones / Q000378:metabolism; D015394:Molecular Structure; D009690:Nucleic Acid Conformation; D009707:Nucleosomes / Q000235:genetics* / Q000378:metabolism; D011743:Pyrimidines / Q000032:analysis; D014466:Ultraviolet Rays
D002843:Chromatin; D006657:Histones; D009707:Nucleosomes; D011743:Pyrimidines; D004247:DNA; C030986:pyrimidine