Selective exposure of antigenic determinants in chromosomal proteins upon gene activation in polytene chromosomes.

The immunological accessibility of the nucleosomal core protein H3 and non-histone protein, HMG-1, was studied in transcriptionally active regions of Chironomus thummi polytene chromosomes. Chromosomal loci were decondensed by incubating isolated salivary glands in various salt solutions or hemolymph. Indirect immunofluorescence studies on these chromosomes using anti-sera to histone H3 revealed that the puffed regions were depleted of fluorescence. The lack of fluorescence could be correlated with the degree of puffing and the level of transcriptional activity. The puffed regions fluoresce after anti-H3 addition if the chromosomes are not cross-linked with formaldehyde, and if prior to the addition of antibodies the chromosomes are treated with 45% acetic acid. We conclude that, whereas histone H3 is present in the puffed regions, its antigenic determinants are sterically hindered by components which are extractable by 45% acetic acid. On the other hand, the antigenic determinants of protein HMG-1 are always available to antibody binding in puffed regions, as well as other chromosomal areas.