Structural paradox of polytene chromosomes.

The observation of thick chromatin fibers in interbands of Dipteran polytene chromosomes suggests that there should be 5 to 10 times more mass and DNA in interbands than is commonly thought to be present. To resolve this paradox, the chromatin content of interbands was estimated, using whole-mounted polytene chromosomes from Drosophila melanogster. Densitometry of high voltage electron microscopic negatives provides an estimate of less than 4:1 for the average ratio of cross-sectional dry mass (or mass per unit chromosome length) of bands relative to interbands. This ratio, combined with an estimate of the length of chromosome composed of interbands, indicates that at least 26% of chromosome mass is contributed by interband chromatin. Since DNA comprises a similar proportion of chromatin mass in bands and interbands (Laird et al., 1980b), these data imply that DNA sequences in interbands represent at least 26% of the euchromatic genome of D. melanogaster. This result calls for reinterpretation of some of the genetic and molecular data from Diptera. The discrepancy between this higher estimate of interband mass and DNA, and previous estimates of 3-5%, is discussed. One possibility is that previous measurements were made on prominent interbands, which are proposed here to be in regions that are delayed in DNA replication. Such interbands would be reduced in polyteny and DNA content compared with the average interband region. The concept of local variations in polyteny is also used here to explain major differences in the cross-sectional mass of bands. This leads to a revised model of polytene chromosomes in which at least three levels of polyteny, rather than one or two levels, can be present within one euchromatic region.