A family of unusually spliced biologically active transcripts encoded by a Drosophila clock gene.

Complementary DNA cloning of the transcripts of the Drosophila clock gene period reveals three distinct transcripts. These result from unusual splicing pathways, one involving a CG 3' splice site and one resulting in the use of two different reading frames in one exon, and they predict three separate proteins. Two of the cloned cDNAs can restore clock function to mutant arrhythmic flies.

Molecular mapping of point mutations in the period gene that stop or speed up biological clocks in Drosophila melanogaster.

The pero1 and the pers mutations in Drosophila melanogaster, which seem to eliminate or speed up, respectively, the clocks underlying biological rhythmicity, were mapped to single nucleotides. Chimeric DNA fragments consisting of well-defined wild-type plus mutant DNA subsegments were constructed, introduced into flies by germ-line transformation, and assayed for biological activity. These experiments localized both pero1 and pers to a 1.7-kilobase DNA fragment that is mostly coding DNA. Sequencing of this subsegment from each mutant showed that pero1 is completely accounted for by a nonsense mutation in the third coding exon of a 4.5-kilobase RNA transcribed from this locus. The pers mutation is also a single nucleotide substitution, in the fourth coding exon, which results in a serine-to-asparagine substitution in the per gene protein product. The functional significance of these changes is discussed with reference to the phenotypes of the two mutations.

The period clock locus of D. melanogaster codes for a proteoglycan.

The period (per) gene of D. melanogaster is involved in the generation of biological rhythms. The most striking feature of the predicted coding sequence, corresponding to the key 4.5 kb transcript from this locus, is an extensive run of alternating Gly-Thr residues. This is homologous to a series of Gly-Ser repeats in a chondroitin sulfate proteoglycan. To determine whether the per transcript codes for a proteoglycan, a region of its coding sequence was expressed (in bacteria) as part of a fusion protein, which was used to immunize rabbits. When the resultant immune sera were used to probe fly protein preparations, they detected an antigen that is present in wild-type flies and absent in a per- mutant. Biochemical characterization of this antigen indicated that it is indeed a proteoglycan.