Nucleic Acids Research, 1994, Vol. 22, No. 23 5068-5075
© 1994
MOLECULAR BIOLOGY |
Unusual properties of genomic DNA molecules spanning the euchromatic – heterochromatic junction of a Drosophila minichromosome
Laboratory of Developmental Genetics, Wadsworth Center New York State Department of Health, PO Box 22002, Albany, NY 12201-2002 1Howard Hughes Medical Institute Research Laboratories, Carnegie Institution of Washington, Department of Embryology 115 West University Parkway, Baltimore, MD 21210, USA
*To whom correspondence should be addressed
Received June 28, 1994. Revised September 29, 1994. Accepted September 29, 1994.
While investigating the copy number of minichromo-some Dp(1;f) 1187 sequences in the polyploid chromo-somes of ovarian nurse and follicle cells of Drosophila melanogaster we discovered that restriction fragments spanning the euchromatic – heterochromatic junction of the chromosome and extending into peri-centro-meric sequences had the unusual property of being selectively resistant to transfer out of agarose gels during Southern blotting, leading to systematic reduc-tions in Dp 1187-specific hybridization signals. This property originated from the peri-centromeric sequences contained on the junction fragments and was persistently associated with Dp1187 DNA, despite attempts to ameliorate the effect by altering experi-mental protocols. Transfer inhibition was unlikely to be caused by an inherent physical property of repetitive DNA sequences since, in contrast to genomic DNA, cloned restriction fragments spanning the euchro-matic – heterochromatic junction and containing repetitive sequences transferred normally. Finally, the degree of inhibition could be suppressed by the addition of a Y chromosome to the genotype. On the basis of these observations and the fact that peri-centromeric regions of most eukaryotic chromosomes are associated with cytologically and genetically defined heterochromatin, we propose that peri-centro-meric sequences of Dpi 187 that are incorporated into heterochromatin in vivo retain some component of heterochromatic structure during DNA isolation, perhaps a tightly bound protein or DNA modification, which subsequently causes the unorthodox properties observed in vitro.
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