Each element of theDrosophilatRNAArggene split promoter directs transcription inXenopusoocytes

Sharp, S; Dingermann, Theo; Schaack, Jerome; Sharp, J A; Burke, David; Derobertis, E; Söll, Dieter

doi:10.1093/nar/11.24.8677

Cited by 16 publications

(13 citation statements)

References 39 publications

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“…This suggests that the control region most important for initiation is the D-control region as suggested for a C. elegans tRNAPro gene (5) and a Drosophila tRNAArg gene (40).…”

Section: Resultsmentioning

confidence: 90%

“…The inside initiation site appears to be chosen by a mechanism that resembles measuring back from an internal control region (5,40). It remains relatively constant with respect to the mature coding sequence at a position that appears to be characteristic for the tRNAHis gene.…”

Section: Resultsmentioning

confidence: 99%

“…The formation of eucaryotic tRNA precursors by RNA polymerase III and specific transcription factors (3,15,31,36,41) is regulated by two internal control regions (4,6,14,16,20,38,40). Transcription of tRNA genes is also affected by elements in the 5'-flanking sequence.…”

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confidence: 99%

“…The results of a recent study on the in vivo transcription of Xenopus oocytes of 5' deletion mutants of a Drosophila tRNAArg gene suggest that a measurement function is also responsible for the selection of the initiation nucleotide in tRNA gene transcription (40). The stringent selection of the initiation site is altered when part or all of the D-control region is deleted.…”

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confidence: 99%

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Transcription factor binding is limited by the 5'-flanking regions of a Drosophila tRNAHis gene and a tRNAHis pseudogene.

Cooley¹,

Schaack²,

Burke³

et al. 1984

Mol. Cell. Biol.

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We determined the sequence of a Drosophila tRNA gene cluster containing a tRNAHis gene and a tRNAHis pseudogene in close proximity on the same DNA strand. The pseudogene contains eight consecutive base pairs different from the region of the bona fide gene which codes for the 3' portion of the anticodon stem of tRNAHis. The tRNAHis gene is transcribed efficiently in Drosophila Kc cell extract, whereas the pseudogene is not. The pseudogene is also a much poorer competitor than the real gene in a stable transcription complex formation assay, even though the sequence alteration in the pseudogene does not affect the sequence or spacing of the putative internal transcription control regions. Recombinant clones were constructed in which the 5'-flanking regions are exchanged. The transcription efficiencies and competitive abilities of the recombinant clones resemble those of the genes from which the 5' flank was derived; for example, the tRNAHis pseudogene with the 5'-flanking sequence of the tRNAHis gene is now efficiently transcribed. Deletion analysis of the pseudogene 5' flank failed to uncover an inhibitory element. Deletion analysis of the real gene showed very high dependence on the presence of the wild-type 5'-flanking sequence for factor binding to the internal control regions and stable complex formation. The 5'-flanking sequence of a Drosophila tRNAArg gene active in the Drosophila Kc cell extract does not restore transcriptional activity or stable complex formation. The tRNAHis gene and pseudogene behave atypically in HeLa cell extract. Both genes compete for HeLa transcription factors, but neither of them is efficiently transcribed. Removal of the 5'-flanking sequences of each gene and replacement with various sequences, including the tRNAArg gene 5' flank, does not allow increased transcription in HeLa cell extract.

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“…This suggests that the control region most important for initiation is the D-control region as suggested for a C. elegans tRNAPro gene (5) and a Drosophila tRNAArg gene (40).…”

Section: Resultsmentioning

confidence: 90%

Section: Resultsmentioning

confidence: 99%

mentioning

confidence: 99%

mentioning

confidence: 99%

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Transcription factor binding is limited by the 5'-flanking regions of a Drosophila tRNAHis gene and a tRNAHis pseudogene.

Cooley¹,

Schaack²,

Burke³

et al. 1984

Mol. Cell. Biol.

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“…in some cases it appears that regulatory mechanisms exist in vivo which are not reflected in vitro [31]. One example is the variation between rates of synthesis for the tRNAVa' genes in vitro and in vivo described by Leung et al [32].…”

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confidence: 99%

Steady‐state kinetic analysis of transcription of cloned tRNA^Ser genes from Drosophila melanogaster

Louis

Spiegelman

1985

European Journal of Biochemistry

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Drosophila rnelunoguster Schneider I1 cells contain a factor which inhibits transcription in vitro of cloned tRNA genes in crude extracts made from these cells. The inhibitor could, however, be effectively neutralized by addition of certain non-template DNAs.In the absence of the transcription inhibitor activity, the steady-state kinetics of tRNA production from cloned genes followed one-substrate enzyme kinetics to a high degree of accuracy. Maximal rates of transcription and apparent affinity constants were analyzed for a collection of cloned D. melunogaster tRNASe' genes. The stability of the complex formed by the transcription proteins and the template DNA was found to be nearly constant for the genes examined. The transcription rates, however, were greatly influenced by the DNA sequences flanking the tRNA genes.Analysis of transcription competition between DNA templates showed pure competitive behavior. inhibition constants derived from these experiments indicated that the formation of the transcription complex was affected by sequences flanking the tRNA genes. Furthermore, the rate-limiting step in complex formation was independent of the stability of the final form of the complex. Transcription experiments in vitro have shown that specific transcription requires protein factors which do not normally copurify with RNA polymerase 111 [19,25 -281. Furthermore, during transcription a stable association between the DNA template and at least one of the transcription factors is formed [7,21,29, 301. This formation of a complex can be observed by a time course of synthesis experiment, which shows a lag period in product formation [7, 301 and in competition experiments in which the association between factors and the template leads to exclusion of transcription of a second template added at a later time [7,10,29, 301. The rates of RNA synthesis are usually linear after the lag period for up to 2 h. Transcription reactions in vitro have been used to examine the effects of alterations in DNA sequence on transcription rates and to assess the importance of sequence and structural features of the genes and their variants.

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The 5 S Ribosomal and Other Small RNAs

Dillon

1987

The Gene

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Each element of theDrosophilatRNA^Arggene split promoter directs transcription inXenopusoocytes

Cited by 16 publications

References 39 publications

Transcription factor binding is limited by the 5'-flanking regions of a Drosophila tRNAHis gene and a tRNAHis pseudogene.

Transcription factor binding is limited by the 5'-flanking regions of a Drosophila tRNAHis gene and a tRNAHis pseudogene.

Steady‐state kinetic analysis of transcription of cloned tRNA^Ser genes from Drosophila melanogaster

The 5 S Ribosomal and Other Small RNAs

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Each element of theDrosophilatRNAArggene split promoter directs transcription inXenopusoocytes

Cited by 16 publications

References 39 publications

Transcription factor binding is limited by the 5'-flanking regions of a Drosophila tRNAHis gene and a tRNAHis pseudogene.

Transcription factor binding is limited by the 5'-flanking regions of a Drosophila tRNAHis gene and a tRNAHis pseudogene.

Steady‐state kinetic analysis of transcription of cloned tRNASer genes from Drosophila melanogaster

The 5 S Ribosomal and Other Small RNAs

Contact Info

Product

Resources

About

Each element of theDrosophilatRNA^Arggene split promoter directs transcription inXenopusoocytes

Steady‐state kinetic analysis of transcription of cloned tRNA^Ser genes from Drosophila melanogaster