RNA metabolism during puff induction in Drosophila melanogaster

Ellgaard, Erik G.; Clever, Ulrich

doi:10.1007/bf00326422

Cited by 60 publications

(23 citation statements)

References 28 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…was also examined. In this case, treatment with 100 ,uM sodium arsenite (lane 13) did not result in readthrough. although the HSP70 protein was induced to the same extent as were the heat-shocked samples (data not shown).…”

Section: Methodsmentioning

confidence: 74%

“…Others have reported that heat shock primarily inhibits rRNA processing but has little or no effect on the amount of newly synthesized pre-rRNAs in human cells (47) and cells derived from lower organisms (6,13,27). When studied, the slight decrease in accumulation has appeared to be due primarily to an increased rate of turnover of these rRNAs (6).…”

Section: Discussionmentioning

confidence: 99%

See 1 more Smart Citation

Analysis of pre-rRNAs in heat-shocked HeLa cells allows identification of the upstream termination site of human polymerase I transcription.

Parker¹,

Bond²

1989

Mol. Cell. Biol.

View full text Add to dashboard Cite

Human rRNA precursors from normal or stressed HeLa cells were studied by Si nuclease mapping of unlabeled RNA and by antisense RNase mapping of RNA from cells that had been labeled in vivo with[32p]PO4. Heating cells to 43°C decreased the amount of newly synthesized rRNA to less than 5% of the control level and led to greater than 95% inhibition of transcription termination at a region 355 to 362 nucleotides downstream of the 3' end of 28S rRNA, with readthrough continuing into the next transcription unit. Heating of cells to 42°C led to 60% inhibition of termination at this site; 50% of transcripts that extended into the nontranscribed spacer ended in a region 200 to 210 nucleotides upstream of the polymerase I (Pol I) initiation site. This is presumed to be the human upstream transcription termination site because of the absence of RNAs with a 5' end corresponding to this region, the location relative to the Pol I initiation site (which is similar to the location of upstream terminators in other species), and the fact that it is 15 to 25 nucleotides upstream of the sequence GGGTTGACC, which has an 8-of-9 base identity with the sequence 3' of the downstream termination site. Surprisingly, treatment of cells with sodium arsenite, which also leads to the induction of a stress response, did not inhibit termination. Pol I initiation was decreased to the same extent as termination, which lends support to the hypothesis that termination and initiation are coupled. Although termination was almost completely inhibited at 43°C, the majority of the recently synthesized rRNAs were processed to have the correct 3' end of 28S. This finding suggests that 3'-end formation can involve an endonucleolytic cut and is not solely dependent on exonucleolytic trimming of correctly terminated rRNAs.

show abstract

Section: Methodsmentioning

confidence: 74%

Section: Discussionmentioning

confidence: 99%

Analysis of pre-rRNAs in heat-shocked HeLa cells allows identification of the upstream termination site of human polymerase I transcription.

Parker¹,

Bond²

1989

Mol. Cell. Biol.

View full text Add to dashboard Cite

show abstract

“…include: recovery from prolonged anaerobiosis [e.g., 2 hr under nitrogen (13)], treatment of isolated glands with uncouplers of oxidative phosphorylation [e.g., 2,4-dinitrophenol (14,15)], and treatment of glands with inhibitors of electron transport [e.g., rotenone (4)]. …”

Section: Resultsmentioning

confidence: 99%

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

Lewis¹,

Helmsing²,

Ashburner³

1975

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

161

View full text Add to dashboard Cite

The The pattern of puffing activity in the polytene chromosomes of Drosophila, and other Diptera, changes in a complex way during development. During late larval and "prepupal" development, at least, these changes are under hormonal control (1, 2). Puffing activity can also be controlled by certain environmental treatments; in particular a brief exposure of salivary glands, either in the intact animal or in culture, to high temperature induces specific puffs (3-5).We report the changes in salivary gland protein synthesis that result from temperature shock and the correlated changes in puffing activity. They will enable a formal proof of the hypothesis that puffs are active gene loci (6), a proof that demands the genetic mapping of a putative puff product to the puff itself and the demonstration that mutations which specifically result in the loss of the puffs also result in the absence of its putative product. Preparation of Samples. At the end of the labeling period the radioactive medium was removed with a fine-tipped glass pipette and the glands were flooded with medium containing a 100-fold excess of nonradioactive methionine. If necessary they were then stored at -70°to await further processing. After the wash in nonradioactive medium the glands were washed in cold 10% trichloroacetic acid and then transferred into small glass test tubes. Then they were washed with 10% cold trichloroacetic acid (20 min Control Experiments. The extraction procedure we used is essentially that of Tissieres et al. (9). We have varied the extraction method (for example by using boiling 10% trichloroacetic acid or prolonged chloroform:methanol extraction at 60°) without affecting the results. We have also studied the possible effects of sodium dodecyl sulfate on protease activity by treatment of samples with either phenyl methyl sulfonyl fluoride (10) or L-1-tosylamido-2-phenylethylchlo-3604

show abstract

“…Heat-shock treatments have been found to affect the synthetic activities of Drosophila larvae (13,14,21) and tissue culture cells (22,23). Those studies have shown that heat-induced loci are sites of active RNA synthesis.…”

mentioning

confidence: 99%

“…Other chromosomal loci also showed marked fluorescence; however, 93D was consistently the most intensely fluorescent locus within a particular nucleus. Autoradiographic studies (21,24) have demonstrated enhanced incorporation of labeled uridine into RNA at specific loci in response to heat-shock. Traditionally, acetic acid fixation has been used as a means of preparing chromosome spreads suitable for autoradiographic analysis.…”

mentioning

confidence: 99%

Association of cyclic GMP with gene expression of polytene chromosomes of Drosophila melanogaster.

Spruill¹,

Hurwitz²,

Lucchesi³

et al. 1978

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

View full text Add to dashboard Cite

The distribution of cyclic nucleotides on polytene chromosomes isolated from Drosophila melanogaster salivary glands was examined by using an indirect immunofluorescent technique. With a fixative that minimized the loss of chromosomal proteins, cyclic GMP, but not cyclic AMP, was observed distributed along the chromosomes. The subchromosomal distribution of cyclic GMP correlated with genetically active sites on the chromosomes. After heat-shock treatments, the intensity of cyclic GMP fluorescence was markedly enhanced at specific loci on the chromosomes, with locus 93D as the most intensely fluorescent. Autoradiographic analysis with [3Hjuridine revealed that 93D was the most transcriptionally active locus within a particular nucleus. These observations suggest that cyclic GMP may participate in processes associated with transcription on polytene chromosomes. The involvement of cyclic GMP in nuclear events associated with gene expression is discussed.Cyclic nucleotides have been implicated as intracellular mediators during processes associated with growth in mammalian tissues and animal cells in culture (1, 2). Utilizing an immunocytochemical approach (3, 4), we have examined the distribution of adenosine 3':5'-cyclic monophosphate (cAMP) and guanosine 3':5'-cyclic monophosphate'(cGMP) during growth and/or differentiation in several biological systems (5-7). The consistent localization of cGMP fluorescence in association with nuclear elements in several tissues (5-9) suggested that cGMP was involved in nuclear events in these cells. Of particular interest was the observation of cGMP along the autosomal bivalents of pachytene spermatocytes during the meiotic prophase in rat testis (6,8). Autoradiographic studies in mouse testis (10) have demonstrated a peak of RNA synthesis during the middle pachytene stage. These observations suggested that cGMP might be involved in processes associated with RNA synthesis in these cells.In order to assess further the possible involvement of cyclic nucleotides with transcriptive activities, we have utilized polytene chromosomes from Drosophila salivary glands, a valuable system for studying transcription. The distribution of cyclic nucleotides on polytene chromosomes was examined immunocytochemically to determine if changes in the distribution occurred concurrently with changes in genetic activities at specific loci. Because chromosomal puffing at specific loci is observed during normal development (11) and can be induced by hormone (12) or environmental stimuli (13,14), this system offers the advantage of being able to visualize changes in gene activity (i.e., puffing) under various conditions.In several recent reports, immunocytochemical methods were presented for determining the distribution of chromosomal proteins (15-17), including RNA polymerase 11 (17), on Drosophila polytene chromosomes. We adapted these methods to the localization of cyclic nucleotides on polytene chromosomes. cGMP fluorescence was observed to be distributed along the chromosome arms whereas cAMP...

show abstract

RNA metabolism during puff induction in Drosophila melanogaster

Cited by 60 publications

References 28 publications

Analysis of pre-rRNAs in heat-shocked HeLa cells allows identification of the upstream termination site of human polymerase I transcription.

Analysis of pre-rRNAs in heat-shocked HeLa cells allows identification of the upstream termination site of human polymerase I transcription.

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

Association of cyclic GMP with gene expression of polytene chromosomes of Drosophila melanogaster.

Contact Info

Product

Resources

About