In vitro premature termination in SV40 late transcription.

Pfeiffer, P.; Hay, Nissim; Pruzan, Ronald; Jakobovits, Edward B.; Aloni, Yosef

doi:10.1002/j.1460-2075.1983.tb01403.x

Cited by 30 publications

(12 citation statements)

References 55 publications

(29 reference statements)

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“…Furthermore, some Burkitt's lymphoma cell lines that have deregulated c-myc gene expression also have a concentration of mutations in the first exon and intron that correlate with abrogation of the transcriptional block (8). Similar intragenic transcriptional blocks have been observed in the human histone H3.3 gene (46), the human immunoglobulin ,u heavy-chain gene (19,35), the murine c-myc gene (40), the murine c-myb gene (2), the hamster c-fos gene (17), the Drosophila hsp70 gene (21), the simian virus 40 late region (23,24,25,42), the adenovirus major late transcription unit (16,18,33,39,41), and the human immunodeficiency virus type 1 long terminal repeat (31). Modulation of transcription elongation and termination is therefore a general mechanism for the regulation of gene expression in eucaryotic organisms.…”

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confidence: 64%

Intrinsic sites of transcription termination and pausing in the c-myc gene.

Kerppola¹,

Kane²

1988

Mol. Cell. Biol.

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We have studied transcription elongation and termination in the human c-myc gene. Transcription of c-myc gene sequences with purified mammalian RNA polymerase II revealed several sites of transcription termination and pausing in the vicinity of the exon 1-intron 1 junction. This region previously has been shown to block transcription elongation in vivo by nuclear run-on analysis (D. Bentley and M. Groudine, Nature [London] 321:702-706, 1986). These sites were recognized by purified RNA polymerase II, and we therefore designated them intrinsic sites of termination and pausing. Two of these sites cause termination of RNA polymerase Ill transcription as well. RNA polymerase II terminated transcription in a cluster of seven consecutive T residues in the nontranscribed strand and paused during transcription at three additional sites in this region. The intrinsic sites of transcription termination and pausing described here correspond closely to the 3' ends of transcripts synthesized in Xenopus oocytes injected with plasmids containing the c-myc termination region (D. Bentley and M. Groudine, Cell 53:245-256, 1988). This correspondence suggests that the intrinsic recognition of these termination and pause sites by purified RNA polymerase II may play a role in the transcription elongation block observed in vivo.

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confidence: 64%

Intrinsic sites of transcription termination and pausing in the c-myc gene.

Kerppola¹,

Kane²

1988

Mol. Cell. Biol.

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“…We have recently shown that pulse-labeling with (a-32P) UTP of SV40 transcriptional complexes (VTC), viral minichromosomes, nuclear matrices or isolated nuclei of SV40-infected cells leads to the production of 94-98 nt attenuator RNA (10,11,(16)(17)(18). Fig.…”

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confidence: 99%

Attenuation in SV40 as a mechanism of transcription-termination by RNA polymerase B

Hay

Aloni

1984

Nucl Acids Res

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Nuclei which were isolated from SV40 infected cells with a hypotonic detergent-free buffer were used to establish in vitro conditions which lead to transcription-termination at the attenuation site of SV40. This system allowed us to identify regulatory elements involved in transcription-termination by RNA polymerase B transcribing SV40. Transcription-termination at the attenuation site was found to be ionic strength dependent. Efficient termination occurred at low (100 mM NaCl) but not at high (100 mM (NH4)2 SO4 or 300 mM NaCl) ionic strength. When nuclei were prewashed with 300 mM NaCl, the efficiency of transcription-termination was low even when transcription was carried out at low ionic strength (100 mM NaCl). Efficient transcription-termination in the high salt prewashed nuclei was reconstituted by complementation with a high salt (300 mM NaCl) soluble factor extracted from nuclei of uninfected cells. In addition, the efficiency of transcription-termination was significantly reduced when ITP replaced GTP in the transcription reaction mixture. Our data indicate that a nuclear factor and RNA secondary structure are essential regulatory elements involved in transcription-termination by RNA polymerase B.

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“…In a series of studies with simian virus 40 (SV40) as the experimental system, it has recently been demonstrated that the eucaryotic polymerase B that transcribes SV40 RNA can respond to transcription termination and pausing signals similar to those of the procaryotic polymerase (31,55,56) and that a mechanism resembling attenuation in procaryotes regulates late transcription in SV40 (2,3,31,37,45,55,56). Moreover, a model in which attenuation and modulation of mRNA secondary structure in a feedback control mechanism quantitatively regulate SV40 gene expression has been presented (2,31).…”

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Transcription of minute virus of mice, an autonomous parvovirus, may be regulated by attenuation

Ben-Asher¹,

Aloni²

1984

J Virol

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To characterize the transcriptional organization and regulation of minute virus of mice, an autonomous parvovirus, viral transcriptional complexes were isolated and cleaved with restriction enzymes. The in vivo preinitiated nascent RNA was elongated in vitro in the presence of [ot-32P]UTP to generate runoff transcripts. The lengths of the runoff transcripts were analyzed by gel electrophoresis under denaturing conditions. On the basis of the map locations of the restriction sites and the lengths of the runoff transcripts, the in vivo initiation sites were determined. Two major initiation sites having similar activities were thus identified at residues 201 ± 5 and 2005 ± 5; both of them were preceded by a TATAA sequence. When uncleaved viral transcriptional complexes or isolated nuclei were incubated in vitro in the presence of [a-32P]UTP or [0t-32P]CTP, they synthesized labeled RNA that, as determined by polyacrylamide gel electrophoresis, contained a major band of 142 nucleotides. The RNA of the major band was mapped between the initiation site at residue 201 ± 5 and residue 342. We noticed the potential of forming two mutually exclusive stem-and-loop structures in the 142nucleotide RNA; one of them is followed by a string of uridylic acid residues typical of a procaryotic transcription termination signal. We propose that, as in the transcription of simian virus 40, RNA transcription in minute virus of mice may be regulated by attenuation and may involve eucaryotic polymerase B, which can respond to a transcription termination signal similar to that of the procaryotic polymerase.

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In vitro premature termination in SV40 late transcription.

Cited by 30 publications

References 55 publications

Intrinsic sites of transcription termination and pausing in the c-myc gene.

Intrinsic sites of transcription termination and pausing in the c-myc gene.

Attenuation in SV40 as a mechanism of transcription-termination by RNA polymerase B

Transcription of minute virus of mice, an autonomous parvovirus, may be regulated by attenuation

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