Protein Synthesis In Salivary Glands Research Articles

Ecdysterone induced protein synthesis in salivary glands and in fat body of Drosophila melanogaster larvae.

Salivary glands of 3rd instar larvae of Drosophila melanogaster were labeled with 3H-leucine in the presence and absence of ecdysterone. Twenty-six ecdysterone inducible proteins were detected. Their induction was correlated with puff stage. Synthesis of fifteen proteins commenced during early puff stage (PS2); synthesis of seven others at late puff stages (PS8-10). Synthesis of four proteins was induced between puff stage 3/4 and 7/8. Thus, the hormonal induction of protein synthesis generally reflected the appearance of early and of late puffs as described by Ashburner (1972). Eleven ecdysterone inducible proteins were detected in larval fat body in vitro. Comparison of the fat body to the salivary gland proteins revealed that one of the ecdysterone induced fat body proteins was identical in molecular weight and charge to one of the proteins induced by ecdysterone in salivary glands.

Protein synthesis in salivary glands of Drosophila hydei after experimental gene induction

Several treatments, namely incubation at 37 degrees C, in the presence of arsenite, 2,4-dinitrophenol or vitamin B-6, or release from anaerobiosis induce the same set of puffs in the polythene chromosomes of salivary glands of Drosophila hydei. Analysis of changes in protein-synthetic patterns (as determined by radioautography of sodium dodecyl sulphate-gel electrophoretograms of extracts from [35S]methionine-labelled salivary glands) showed that concomitant with puff induction by these various treatments the same six strongly labelled polypeptide bands appeared. The amount of radioactive label in these peptides accounted for 25% of the total incorporation of [35S]methionine, except during incubation at 37 degrees C when it accounted for about 50%. The rate of synthesis of these peptides was maximal 1 h after the start of the puff-inducing treatment. The rate of decay of the rate of synthesis showed first-order kinetics both after removal of the puff-inducing stimulus or in the presence of actinomycin, with a half-life of approx. 4h.

Parallel changes in puffing activity and patterns of protein synthesis in salivary glands of Drosophila.

The changes in protein synthesis of salivary glands of Drosophila resulting from a brief exposure at 37 degrees have been analyzed on sodium dodecyl sulfate--acrylamide gels. In D. melanogaster and D. hydei this treatment induces nine and six new puffs, respectively, in the polytene chromosomes. After 20 min treatment seven new proteins are synthesized by the glands of D. melanogaster and six by those of D. hydei as detected by [35S]methionine labeling. Other agents, e.g., recovery from anaerobiosis, induce the same puffs and the same proteins. The extent of protein induction and the degree of puff induction are related to the severity of the temperature treatment. The new proteins are detected after 10 min treatmene at 37 degrees and their synthesis is inhibited by actinomycin D. Actinomycin D added 5 min after the start of temperature treatment has little effect on subsequent protein synthesis. The induced proteins are not tissue specific. Electrophoretic differences of two proteins exist between D. melanogaster and D. simulans, encouraging attempts to map the proteins' gene loci and to test directly whether or not the puffs code for them.

Protein synthesis in salivary glands of Drosophila melanogaster: Relation to chromosome puffs

The salivary glands and other tissues from Drosophila melanogaster were dissected at various times throughout the prepupal period, as well as after heat shocks and ecdysterone treatments, and the proteins labelled by incubating the isolated tissues with [ 35S]methionine were separated by electrophoresis on sodium dodecyl sulphate-polyacrylamide gel. The labelled band patterns from salivary gland, as seen on the autoradiograph of the gel, showed striking variations, in a manner remarkably similar to variations in puff patterns during the same prepupal period. In proteins from Malpighian tubes, the pattern of bands varied to a lesser extent and in brain only a few components were modified. Heat shock brought about the appearance of a number of new bands, while others were reduced in intensity. This effect was observed with all the tissues examined, salivary glands, brain and Malpighian tubes, as well as wing imaginal discs, tissue lacking polytene chromosomes. The six most heavily labelled bands induced by heat shock represent about 30%, and one component alone represents over 15%, of the total label in the sample, as seen in salivary glands, brain and Malpighian tubes. The synthesis of RNA at puff sites was investigated after heat shock by [ 3H]uridine labelling. By correlating the amount of [ 3H]uridine in some puffs with the level of [ 35S]methionine in some bands a tentative relation is suggested in a few instances. The effect of ecdysterone treatment was also studied in the salivary glands. Changes in a number of protein bands were noticed, though they were much less pronounced than those following heat shock.