Developmental protein synthesis is required for the transcription of Dictyostelium prespore genes.
It has been established previously that the maintenance of expression of prespore-specific genes of Dictyostelium discoideum is prevented by the translational inhibitor cycloheximide. The drug had no effect upon the level of transcripts of the other genes examined, prestalk-specific or cell type-nonspecific. However, the interpretation of this result is open to question, because of possible nonspecific effects of cycloheximide. We have now characterized the cellular specificity and temporal profiles of mRNA accumulation of additional Dictyostelium cDNA clones, and have examined other inhibitors of in vivo protein synthesis. Four structurally and mechanistically distinct translational inhibitors each prevented the reaccumulation of prespore transcripts in cyclic AMP-primed, disaggregated amoebae. These results establish the importance of developmental protein synthesis in the accumulation of prespore gene transcripts. Nuclear run-on transcription assays were used to learn whether protein synthesis is required primarily for mRNA synthesis or transcript stability. A transcriptional time course first demonstrated that the abundance of these cell-specific transcripts during development mirrors their rates of synthesis. Significantly, the protein synthesis requirement of the prespore genes examined also occurs at the level of mRNA transcription, implying the existence of one or more developmentally regulated transcriptional activators.
Benedict
M A
MA
Department of Biology, Amherst College, MA 01002.
Desilver
D A
DA
Pelletier
D E
DE
Pentz
W H
WH
Ratner
D I
DI
D003513:Cycloheximide / Q000494:pharmacology; D004023:Dictyostelium / Q000235:genetics* / Q000254:growth & development / Q000502:physiology; D005656:Fungal Proteins / Q000096:biosynthesis* / Q000235:genetics; D015966:Gene Expression Regulation, Fungal / Q000187:drug effects; D005800:Genes, Fungal; D007700:Kinetics; D009693:Nucleic Acid Hybridization; D014176:Protein Biosynthesis; D012331:RNA, Fungal / Q000096:biosynthesis; D012333:RNA, Messenger / Q000378:metabolism; D013172:Spores, Fungal / Q000235:genetics; D014158:Transcription, Genetic / Q000187:drug effects
D005656:Fungal Proteins; D012331:RNA, Fungal; D012333:RNA, Messenger; D003513:Cycloheximide