Preferential binding of the carcinogen benzo[a]pyrene to proteins of the nuclear matrix

Rat liver nuclei or hepatocytes were incubated with the procarcinogen benzoa]pyrene (Ba]P) and its ultimate carcinogen, anti-benzoa]pyrene-7,8-diol-9,10-epoxide (BPDE). When nuclei were fractionated by mild micrococcal nuclease digestion into different chromatin regions to determine the distribution of covalent binding to proteins, there was a much higher level of Ba]P bound to proteins of the non-released fraction than to those of released mono- and oligonucleosomes. When non-released material was further fractionated with 2 M NaCl, the highest level of Ba]P binding was found in the proteins of the salt-insoluble fraction. Electrophoretic analysis of 3H]Ba]P-modified nuclear proteins revealed radioactive species migrating in the regions of histones H1 and H3, high mobility group (HMG) proteins 1 and 2, and various high mol. wt non-histone proteins. The non-released fraction contained prominent Ba]P-modified species migrating in the position of the lamins, major components of the nuclear matrix. To confirm Ba]P modification of nuclear matrix proteins, following exposure to Ba]P or BPDE, nuclei were fractionated by a different procedure into an active chromatin fraction, a bulk chromatin fraction, a high-salt-extracted chromatin fraction and a nuclear matrix fraction. Proteins of the nuclear matrix bound consistently more Ba]P metabolites than those of bulk chromatin. This was true following exposure to Ba]P or both low and high concentrations of BPDE, in contrast to previous data on damage to nuclear matrix DNA. Proteins of active chromatin bound more carcinogen than bulk chromatin proteins at low concentrations of BPDE, but less than bulk chromatin at higher concentrations, in parallel with previous data on DNA damage in active chromatin. The potential significance of Ba]P binding to nuclear matrix proteins is discussed.