THE DISTRIBUTION OF GENES ON CHROMOSOMES - A CYTOLOGICAL APPROACH

Studies during the last 20 years have shown that the chromosomes of many organisms, especially those of higher vertebrates, consist of a series of segments having different properties. These can be recognized as, for example, G- and R-bands. Recent studies have indicated that genes tend to lie in the R-bands rather than in the G-bands, although the number of genes that has been mapped with high precision is, as yet, only a very small proportion of the total, probably much less than 1%. We have therefore sought to study the distribution of genes on chromosomes using a cytological approach in conjunction with ''universal'' markers for genes. Such markers include mRNA and the gene-rich, G + C-rich H3 fraction of DNA, both of which can be localized using in situ hybridization, and DNase I hypersensitivity, and digestion by restriction enzymes known to show selectivity for the CpG islands associated with active genes, both of which can be detected using in situ nick translation. We have chosen to use the approaches involving in situ nick translation and have shown that the patterns of DNase I hypersensitivity and of CpG islands on human chromosomes show a strict correspondence to R-banding patterns: Deviations from R-banding patterns reported by previous investigators who have made similar studies appear to be attributable to excessive digestion. On the other hand, we have not found the expected differentiation between the active and inactive X chromosomes; this may perhaps be attributable to such factors as the demethylation of some non-island CpGs in the inactive X and the possible alterations of chromatin structure caused by methanol-acetic-acid fixation affecting DNase I hypersensitivity.