We have sequenced the 480 base pair (bp) repeating unit of the 5S RNA genes of the Dipteran fly Calliphora erythrocephala and compared this sequence to the three known 5S RNA gene sequences from the Dipteran Genus Drosophila (1,2). A striking series of five perfectly conserved homologies identically positioned within the 5' flanks of all four Dipteran 5S RNA coding regions has thus been identified. The spacing (12-13 bp) between all of these homologies is typical of A form rather than B form DNA. Given that the eukaryotic 5S RNA gene specific initiation factor TFIIIA (3) is a DNA unwinding protein (4), a role for these Dipteran 5' flank homologies in initiation site selection on 5S RNA genes transiently unwound for transcription is suggested. One of the Dipteran homology blocks is highly conserved in sequence and position in all but one of the eukaryotic 5S RNA gene sequences known to date (17/18 genes). Its sequence (consensus: TATAAG) and position (average center: -26 bp) are highly reminiscent of the polymerase II gene 'TATA' box (5).
In most species of dipteran fly examined, a fraction of the rDNA cistrons are interrupted by introns. These dipteran intron+ rRNA genes are unique in that they are transcriptionally inactive. Previous studies have investigated the mechanism underlying this transcriptional repression for rRNA genes carrying the best characterized sequence family of such introns, the so-called type 1 introns first identified in Drosophila melanogaster. These studies have established that cloned examples of both intron-free and type 1 intron+ rRNA genes will support transcription in a cell-free system and suggest therefore that a difference in the chromatin state of the two gene types must underlie their very different potential for in vivo transcription. We have examined this possibility for the type 1 intron+ rDNA cistrons of Calliphora erythrocephala by in situ hybridization studies using the polytene chromosome complement of the pupal bristle-forming (trichogen) cells. These studies show that the chromatin configuration of the two gene types is strikingly different. The intron-free genes are preferentially localized in the actively transcribed fibrillar center of the nucleolus. The intron+ genes are preferentially condensed in the blocks of heterochromatin attached to the nucleolus.
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