Nucleoprotein localization by bismuth staining.

Experiments on isolated mouse liver muclei involving enzyme digestion, the crosslinking of amino groups and alkaline hydrolysis demonstrate that bismuth binds to nucleoproteins through amino and phosphate groups. Analysis of the nucleoproteins extracted with salt and acid solutions in conjunction with bismuth staining after these treatments suggests that: (1) a bismuth amino group interaction occurs on ribonucleo-protein particles, histones and perhaps some non-histone chromosomal proteins, and (2) bismuth phosphate binding is specific for one, or all, of three distinct species of non-histone proteins. These results suggest that histones not tightly bound to DNA through their amino groups are present on interchromatin granules, the presumed transcriptionally active regions of chromatin. Phosphorylated non-histone proteins are also localized at these sites. Staining with heavy metals such as bismuth may be the best method for high resolution localization of nucleoproteins involved with regulating gene activity and maintaining chromatin structure.