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J Rebers

Publications and source records attributed to J Rebers.

2 recordsLinked to original sources

Overlapping antiparallel transcripts induced by ecdysone in a Drosophila cell line.

Treating Drosophila Kc tissue culture cells with 20-hydroxyecdysone (20E) causes a rapid 10-fold increase in the synthesis of three polypeptides, with molecular weights of 28 kD, 29 kD, and 40 kD (named EIP28, EIP29, and EIP 40, in reference to their molecular weights), as shown by Savakis et al. (1980, 1984). Genomic clones that include Eip40, a gene encoding the EIP40 polypeptide, have been isolated and used to characterize transcription in a 10 kilobase region. At least five different transcripts are produced from this region in Kc cells. One of these transcripts is overlapping and antiparallel to the Eip40 transcript. Although these genes are located quite near one another, their response to 20E varies from little, if any, induction to an induction that is at least 25 fold. Approximately 2.5 kb of DNA which encompasses the Eip40 gene has been sequenced. Analysis of this sequence shows that Eip40 encodes a Drosophila homologue of the enzyme cystathionase. The complementary DNA strand, which produces an 1800 N transcript, has a 645 N open reading frame (ORF). This ORF shows some similarity to cystathionase, the protein encoded by the complementary strand, but shows no detectable similarity to other reported protein sequences.

Amino Acid Sequence↗

Developmental expression of three genes for larval cuticular proteins of the tobacco hornworm, Manduca sexta.

Three cDNA clones coding for the 12.8, 13.3, and 14.6 kDa larval cuticular proteins of the tobacco hornworm, Manduca sexta, were isolated and characterized. Hybridization to abdominal epidermal RNA from different stages showed that the genes for the 12.8 and 13.3 kDa proteins were expressed only during larval life. By contrast, the gene for the 14.6 kDa protein was expressed throughout the segment during the feeding, growing larval stages, then only in the flexible intersegmental regions during the deposition of endocuticle in the pharate pupa and adult. Quantitative RNA dot blot hybridizations showed that the RNA for each protein disappeared during the larval molt when the ecdysteroid titer was high, then reappeared during the preecdysial deposition of endocuticle. All disappeared when the epidermis became pupally committed at the onset of wandering. Exposure of the fourth instar epidermis to 20-hydroxyecdysone (20HE) in vitro under conditions that lead to the formation of a new larval cuticle by 48 hr caused the disappearance of these RNAs by 18 hr. Exposure of Day 2 fifth instar epidermis to 20HE in vitro caused a depression of these RNAs which in the case of the RNAs coding for the 12.8 and 13.3 kDa proteins was partially prevented by simultaneous exposure to methoprene, a juvenile hormone (JH) mimic. By contrast, the RNA for the 14.6 kDa protein was suppressed by exposure to methoprene alone. Thus, each of these larval cuticular genes is turned off by high ecdysteroid; the presence or absence of JH determines whether or not this suppression is permanent in some or all cells.

Animals↗