Late messenger RNA production by viable simian virus 40 mutants with deletions in the leader region

Piatak, Michael; Subramanian, K. N.; Roy, Pranab; Weissman, Sherman M.

doi:10.1016/0022-2836(81)90409-5

Cited by 77 publications

(79 citation statements)

References 45 publications

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“…We regard this region as a strong potential candidate for a DNA replication origin structure. A 17 nucleQtide A + T sequence and several palindromes are present in the region of the SV40 origin of replication (Reddy et al, 1978;Piatak et al, 1981). The tandem reiterations found in the Vmw 12 and Vmw 68 gene introns are a feature which resembles three sets of tandem reiterations present at the other end of R s (Davison & Wilkie, 1981).…”

Section: Discussionmentioning

confidence: 99%

DNA Sequence Anaylysis of an Immediate-Early Gene Region of the Herpes Simplex Virus Type 1 Genome (Map Coordinates 0.950 to 0.978)

Murchie

McGeoch

1982

Journal of General Virology

113

125

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SUMMARYThe nucleotide sequence of 4 kilobases of DNA from within the short region of the genome of herpes simplex virus type 1 has been determined. This portion of DNA contains the junctions of the terminal and internal inverted repeat sequence components with the unique sequence component and the 5' regions of the genes which encode the Vmw 12, Vmw 68 and Vmw 175 immediate-early polypeptides. The transcription and translation initiation sites of all three genes and the 5' and 3' boundaries of the Vmw 12 and Vmw 68 gene introns have been localized on the DNA sequence and shown to be flanked by sequences which resemble those found in similar positions in other eukaryotic genes. For the Vmw 12 and Vmw 68 genes the promoters, the 5'-non-coding regions of the mRNAs, and the introns lie within the terminal and internal inverted repeats respectively while the polypeptide-coding regions lie in the short unique component. The introns consist largely of tandemly reiterated copies of a 22-nucleotide sequence: the Vmw 12 gene intron contains seven of these and the Vmw 68 gene intron five. The Vmw 175 gene is located entirely within the short repeats, of which those regions sequenced here have the extremely high G + C content of 78 %, in marked contrast to the value of 66 % obtained for the adjacent region of the unique sequence component. Prediction of the complete amino acid sequence of the Vmw 12 polypeptide, accounting for a mol. wt. of 9830, and of the first 523 amino-terminal amino acids of the Vmw 175 polypeptide has been made from the DNA sequence. The polypeptide-coding region of the Vmw 175 gene has an average G + C content of 80% but nevertheless specifies a wide range of amino acid types because of the maximal assignment of G and C residues to codon third base positions.

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Section: Discussionmentioning

confidence: 99%

DNA Sequence Anaylysis of an Immediate-Early Gene Region of the Herpes Simplex Virus Type 1 Genome (Map Coordinates 0.950 to 0.978)

Murchie

McGeoch

1982

Journal of General Virology

113

125

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“…This Ara C concentration had been previously determined to be inhibitory to plasmid DNA replication but not lethal to cells during the course of the transfection (24 h). The normal period of 48 h was not used because some cell death could be detected by this time.…”

mentioning

confidence: 99%

“…Total RNA was extracted from cells 48 h after transfection by the method of Villarreal (69). The RNA preparations were freed of DNA by treatment for 45 min at 37°C with RNase-free DNase prepared by described methods (68).…”

mentioning

confidence: 99%

Analysis of an activatable promoter: sequences in the simian virus 40 late promoter required for T-antigen-mediated trans activation.

Keller¹,

Alwine²

1985

Mol. Cell. Biol.

129

138

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The late promoter of simian virus 40 (SV40) is activated in trans by the viral early gene product, T antigen. We inserted the wild-type late-promoter region, and deletion mutants of it, into chloramphenicol acetyltransferase transient expression vectors to identify promoter sequences which are active in the presence of T antigen. We defined two promoter activities. One activity was mediated by a promoter element within simian virus 40 nucleotides 200 to 270. The activity of this element was detectable only in the presence of an intact, functioning origin of replication and accounted for 25 to 35% of the wild-type late-promoter activity in the presence of T antigen. The other activity was mediated by an element located within a 33-base-pair sequence (simian virus nucleotides 168 to 200) which spans the junction of the 72-base-pair repeats. This element functioned in the absence of both the origin of replication and the T-antigen-binding sites and appeared to be responsible for trans-activated gene expression. When inserted into an essentially promoterless plasmid, the 33-base-pair element functioned in an orientation-dependent manner. Under wild-type conditions in the presence of T antigen, the activity of this element accounted for 65 to 75% of the late-promoter activity. The roles of the 33-base-pair element and T antigen in trans-activation are discussed.Transcription appears to be a key point for control of eucaryotic gene expression. Both the utilization of a transcriptional unit and the rate of its transcription can be controlled in response to environmental, metabolic, or developmental signals. Our present understanding of eucaryotic transcriptional control stems, in large part, from studies of the DNA viruses. One specific system, simian virus 40 (SV40), has repeatedly proven to be a useful model of both viral and cellular gene expression mechanisms. The circular, 5,243-base-pair (bp) SV40 genome is relatively simple, containing only two transcription units which are temporally expressed, early and late, during the lytic cycle (66). The early region is transcribed soon after final uncoating in the cell nucleus. The level of transcription from the early promoter is negatively autoregulated by the early gene product, T antigen (4, 34, 41, 51, 53, 61-63, 65). Conversely, late-region transcription is activated by T antigen (11,36,40), and late mRNA accumulates in large quantities in the later phases of the infection, after viral DNA replication (also activated by T antigen) has been initiated.The promoter for early transcription has been studied in detail. In common with many other RNA polymerase II promoters, it contains a Goldberg-Hogness TATA sequence which directs the site of transcription initiation (5,6,24,57). In addition, efficient transcription from the early promoter requires (i) specific binding of cellular factor(s) to guanine * cytosine-rich sequences within the 21-bp repeated region (see Fig. 1 ; 19, 20) and (ii) cis-activation by the 72-bp enhancer elements (22,23,31,43, 71). Any cellular fact...

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“…To overcome these problems, a method was needed that eliminates the heterogeneity at the 3' ends of the cDNAs copied from the various species of mRNA molecules. Piatak et al (14) attempted this by cleavage of the cDNAs with restriction endonucleases that cleave single-stranded DNA. Unfortunately, this procedure is not quantitative because one cannot cleave single-stranded DNAs to completion with restriction endonucleases.…”

Section: Resultsmentioning

confidence: 99%

“…Several groups of workers have analyzed in detail the structures of the viral RNAs synthesized by wildtype (WT) and viable mutants of SV40 having deletions in the late leader region (10,13,14). Based upon these data, it has been hypothesized that the precise sequences at the 5' end of a primary late strand transcript of SV40 might influence the choice of splice sites used in its processing (10,13).…”

Section: Nucleic Acids Researchmentioning

confidence: 99%

Exon mutations that affect the choice of splice sites used in processing the SV40 late transcripts

1985

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ABSTRACrThe spliced species of late SV40 RNAs present in the cytoplasm of cells infected with various wild-type and mutant strains of SV40 that differ in their leader regions were determined using a novel modification of the primer extension method and the S1 nuclease mapping technique. These data indicated that mutations within the first exon of the late RNAs can affect dramatically the utilization of downstream donor and acceptor splice sites. In one instance, a ten base pair insertion within the predominant first exon increased utilization of an infrequently utilized donor splice site such that the small alteration became part of an intervening sequence, thereby suggesting a novel mechanism for regulation of gene expression. In addition, our method enabled detection of a previously unidentified spliced species, representing less than one percent of the SV40 late 19S RNA present in cells infected with wild-type virus, that may be an intermediate in the synthesis of a known doubly spliced 16S RNA species of SV40.

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Late messenger RNA production by viable simian virus 40 mutants with deletions in the leader region

Cited by 77 publications

References 45 publications

DNA Sequence Anaylysis of an Immediate-Early Gene Region of the Herpes Simplex Virus Type 1 Genome (Map Coordinates 0.950 to 0.978)

DNA Sequence Anaylysis of an Immediate-Early Gene Region of the Herpes Simplex Virus Type 1 Genome (Map Coordinates 0.950 to 0.978)

Analysis of an activatable promoter: sequences in the simian virus 40 late promoter required for T-antigen-mediated trans activation.

Exon mutations that affect the choice of splice sites used in processing the SV40 late transcripts

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