Actinomycin and Puromycin: Effects on Sequential Gene Activation by Ecdysone

Clever, Ulrich

doi:10.1126/science.146.3645.794

Cited by 161 publications

(35 citation statements)

References 11 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…This hormone stimulated the incorporation of radioactive uridine into RNA at specific sites on these chromosomes, causing puffing at these loci in a temporarily defined sequence. Furthermore, puromycin was shown to affect "late" puffing but not "early" puffing (23)(24)(25).…”

Section: Discussionmentioning

confidence: 96%

“…Therefore, the existence of a 2-hr delay suggests that there may be intermediate processes between the binding of steroid receptors to chromatin and the induction of this specific mRNA. Evidence for such indirect phenomena is also provided by studies showing that the insect hormone ecdysone causes puffing on the polytenic salivary gland chromosomes of Chironomus (23,24) and Drosophila (25). This hormone stimulated the incorporation of radioactive uridine into RNA at specific sites on these chromosomes, causing puffing at these loci in a temporarily defined sequence.…”

Section: Discussionmentioning

confidence: 98%

See 1 more Smart Citation

Cycloheximide inhibition of hormonal induction of alpha 2u-globulin mRNA.

Chen¹,

Feigelson²

1979

Proc. Natl. Acad. Sci. U.S.A.

View full text Add to dashboard Cite

The induction of hepatic a2u-globulin synthesis by glucocorticoids in isolated hepatocytes occurs via an increase in the level of its mRNA as measured by cell-free translation and by hybridization to an a2u-globulin cDNA probe. To explore whether induction of this mRNA is a direct or an indirect consequence of the interaction of the dexamethasone-receptor complex with the a2u-globulin genome, the requirement for ongoing protein synthesis was examined. Concentrations of cycloheximide too low to prevent precursor incorporation into total poly(A).containing RNA do prevent the hormonal induction of a2u-globulin mRNA. Furthermore, incorporation of 3H-labeled amino acids into total protein was decreased by only 40-50%, and the appearance of the dexamethasone-induced glycosylated forms of a2u-globulin was completely prevented in these cycloheximide-treated hepatocytes. The results suggest that the synthesis of a protein mediator(s) may be required for the induction of a2u-globulin mRNA by glucocorticoids and that the steroid-receptor complex may not interact directly with the a2u-globulin genome. a2u-Globulin is present in the urine of male rats but not in the urine of female rats (1). This protein, which is synthesized in the parenchymal cells of male rat liver and represents approximately 1% of hepatic protein synthesis, is secreted into the serum and excreted in the urine (2-4). In this laboratory, it has been shown that the in vivo regulation of the synthesis of this protein by glucocorticoids (5), thyroid hormones (6), and androgens (7) occurs via modulation of the level of hepatic mRNA coding for ct2u-globulin, whereas pituitary growth hormone stimulates the synthesis of this protein translationally (8).A hepatocyte suspension system that permits the study of hormonal control of (t2u-globulin synthesis under defined conditions in vitro has been developed (9, 10). Isolated hepatocytes synthesize and secrete (a2u-globulin and other hepatic proteins in vitro at an approximately linear rate throughout 30 hr of incubation. Hepatocytes isolated from animals in different endocrine states synthesize a2u-globulin in vitro at rates consistent with the hormonal effects upon its in vivo biosynthesis (11).Increased synthesis of oau-globulin can be induced in vitro by addition of glucocorticoids to hepatocytes derived from male rats in which the rate of synthesis of this protein had been depressed by adrenalectomy, castration, or estrogen pretreatment (11). The induction of (2,,-globulin synthesis by dexamethasone in such hepatocytes is highly specific and occurs by selective increase of aou-globulin mRNA in these cells, as measured by a wheat germ cell-free translational system and by hybridization to a o2,1-globulin cDNA probe (12). The induced intracellular (YoU-globulin or the translation product generated in the wheat germ system using mRNA isolated from the dexaThe publication costs of this article were defrayed in part by page charge payment. This article must therefore be hereby marked "advertisement' in accord...

show abstract

Section: Discussionmentioning

confidence: 96%

Section: Discussionmentioning

confidence: 98%

Cycloheximide inhibition of hormonal induction of alpha 2u-globulin mRNA.

Chen¹,

Feigelson²

1979

Proc. Natl. Acad. Sci. U.S.A.

View full text Add to dashboard Cite

show abstract

“…This sequential appearance has generally been attributed to delayed transcription of some genes relative to others (1)(2)(3). It is thought that genes with compact chromatin or negative histone marks take some time to unravel or to acquire activating histone marks (4)(5)(6)(7)(8).…”

mentioning

confidence: 99%

RNA splicing regulates the temporal order of TNF-induced gene expression

Hao

Baltimore

2013

Proc. Natl. Acad. Sci. U.S.A.

View full text Add to dashboard Cite

When cells are induced to express inflammatory genes by treatment with TNF, the mRNAs for the induced genes appear in three distinct waves, defining gene groups I, II, and III, or early, intermediate, and late genes. To examine the basis for these different kinetic classes, we have developed a PCR-based procedure to distinguish pre-mRNAs from mRNAs. It shows that the three groups initiate transcription virtually simultaneously but that delays in splicing characterize groups II and III. We also examined the elongation times, concluding that pre-mRNA synthesis is coordinate but splicing differences directly regulate the timing of mRNA production.

show abstract

“…Ever since the pioneering work of Clever (1964) and Ashburner (see, e.g., Ashburner et al 1974) on hormone-induced puffing in salivary gland chromosomes suggested that ecdysone might act by regulating transcription, it has been attractive to suppose that all of the steroid hormones are functionally related and that they control transcription by similar mechanisms. If so, studies of ecdysone action in Drosophila should contribute significantly to our understanding of the mechanistic bases of the developmental specificity of steroid effects.…”

mentioning

confidence: 99%