scholarly journals Infectious linear DNA sequences replicating in simian virus 40-infected cells.

1977 ◽  
Vol 21 (2) ◽  
pp. 565-578 ◽  
Author(s):  
P Gruss ◽  
G Sauer
Keyword(s):  
Dna Sequences ◽  
Simian Virus 40 ◽  
Simian Virus ◽  
Linear Dna ◽  
Infected Cells

Circular and linear simian virus 40 DNAs differ in recombination

1985 ◽  
Vol 5 (4) ◽  
pp. 869-880
Author(s):  
D Dorsett ◽  
I Deichaite ◽  
E Winocour
Keyword(s):  
Dna Sequences ◽  
Tandem Repeats ◽  
Regulatory Region ◽  
Simian Virus 40 ◽  
Simian Virus ◽  
T Antigen ◽  
Linear Forms ◽  
Rolling Circle ◽  
Linear Dna ◽  
Sv40 Dna

Linear forms of simian virus 40 (SV40) DNA, when added to transfection mixtures containing circular SV40 and phi X174 RFI DNAs, enhanced the frequency of SV40/phi X174 recombination, as measured by infectious center in situ plaque hybridization in monkey BSC-1 cells. The sequences required for the enhancement of recombination by linear DNA reside within the SV40 replication origin/regulatory region (nucleotides 5,171 to 5,243/0 to 128). Linearization of phi X174 RFI DNA did not increase the recombination frequency. The SV40/phi X174 recombinant structures arising from transfections supplemented with linear forms of origin-containing SV40 DNA contained phi X174 DNA sequences interspersed within tandem head-to-tail repeats derived from the recombination-enhancing linear DNA. Evidence is presented that the tandem repeats are not formed by homologous recombination and that linear forms of SV40 DNA must compete with circular SV40 DNA for the available T antigen to enhance recombination. We propose that the enhancement of recombination by linear SV40 DNA results from the entry of that DNA into a rolling circle type of replication pathway which generates highly recombinogenic intermediates.

scholarly journals Glycosylation and surface expression of the influenza virus neuraminidase requires the N-terminal hydrophobic region.

1984 ◽  
Vol 4 (1) ◽  
pp. 8-16 ◽  
Author(s):  
L Markoff ◽  
B C Lin ◽  
M M Sveda ◽  
C J Lai
Keyword(s):  
Influenza Virus ◽  
Dna Sequences ◽  
Sodium Dodecyl ◽  
Simian Virus 40 ◽  
Simian Virus ◽  
Wild Type ◽  
Hydrophobic Region ◽  
Infected Cells ◽  
Late Region ◽  
Na Gene

A full-length double-stranded DNA copy of an influenza A virus N2 neuraminidase (NA) gene was cloned into the late region of pSV2330, a hybrid expression vector that includes pBR322 plasmid DNA sequences and the simian virus 40 early region and simian virus 40 late region promoters, splice sequences, and transcription termination sites. The protein encoded by the cloned wild-type NA gene was shown to be present in the cytoplasm of fixed cells and at the surface of "live" or unfixed cells by indirect immunofluorescence with N2 monoclonal antibodies. Immunoprecipitation and sodium dodecyl sulfate-polyacrylamide gel electrophoretic analysis of [35S]methionine-labeled proteins from wild-type vector-infected cells with heterospecific N2 antibody showed that the product of the cloned NA DNA comigrated with glycosylated NA from influenza virus-infected cells, remained associated with internal membranes of cells fractionated into membrane and cytoplasmic fractions, and could form an immunoprecipitable dimer. NA enzymatic activity was detectable after simian virus 40 lysis of vector-infected cells. These properties of the product of the cloned wild-type gene were compared with those of the polypeptides produced by three deletion mutant NA DNAs that were also cloned into the late region of the pSV2330 vector. These mutants lacked 7 (dlk), 21 (dlI), or all 23 amino acids (dlZ) of the amino (N)-terminal variable hydrophobic region that anchors the mature wild-type NA tetrameric structure in the infected cell or influenza viral membrane. Comparison of the phenotypes of these mutants showed that this region in the NA molecule also includes sequences that control translocation of the nascent polypeptide into membrane organelles for glycosylation.

Glycosylation and surface expression of the influenza virus neuraminidase requires the N-terminal hydrophobic region

1984 ◽  
Vol 4 (1) ◽  
pp. 8-16
Author(s):  
L Markoff ◽  
B C Lin ◽  
M M Sveda ◽  
C J Lai
Keyword(s):  
Influenza Virus ◽  
Dna Sequences ◽  
Sodium Dodecyl ◽  
Simian Virus 40 ◽  
Simian Virus ◽  
Wild Type ◽  
Hydrophobic Region ◽  
Infected Cells ◽  
Late Region ◽  
Na Gene

A full-length double-stranded DNA copy of an influenza A virus N2 neuraminidase (NA) gene was cloned into the late region of pSV2330, a hybrid expression vector that includes pBR322 plasmid DNA sequences and the simian virus 40 early region and simian virus 40 late region promoters, splice sequences, and transcription termination sites. The protein encoded by the cloned wild-type NA gene was shown to be present in the cytoplasm of fixed cells and at the surface of "live" or unfixed cells by indirect immunofluorescence with N2 monoclonal antibodies. Immunoprecipitation and sodium dodecyl sulfate-polyacrylamide gel electrophoretic analysis of [35S]methionine-labeled proteins from wild-type vector-infected cells with heterospecific N2 antibody showed that the product of the cloned NA DNA comigrated with glycosylated NA from influenza virus-infected cells, remained associated with internal membranes of cells fractionated into membrane and cytoplasmic fractions, and could form an immunoprecipitable dimer. NA enzymatic activity was detectable after simian virus 40 lysis of vector-infected cells. These properties of the product of the cloned wild-type gene were compared with those of the polypeptides produced by three deletion mutant NA DNAs that were also cloned into the late region of the pSV2330 vector. These mutants lacked 7 (dlk), 21 (dlI), or all 23 amino acids (dlZ) of the amino (N)-terminal variable hydrophobic region that anchors the mature wild-type NA tetrameric structure in the infected cell or influenza viral membrane. Comparison of the phenotypes of these mutants showed that this region in the NA molecule also includes sequences that control translocation of the nascent polypeptide into membrane organelles for glycosylation.

scholarly journals Circular and linear simian virus 40 DNAs differ in recombination.

1985 ◽  
Vol 5 (4) ◽  
pp. 869-880 ◽  
Author(s):  
D Dorsett ◽  
I Deichaite ◽  
E Winocour
Keyword(s):  
Dna Sequences ◽  
Tandem Repeats ◽  
Regulatory Region ◽  
Simian Virus 40 ◽  
Simian Virus ◽  
T Antigen ◽  
Linear Forms ◽  
Rolling Circle ◽  
Linear Dna ◽  
Sv40 Dna

Linear forms of simian virus 40 (SV40) DNA, when added to transfection mixtures containing circular SV40 and phi X174 RFI DNAs, enhanced the frequency of SV40/phi X174 recombination, as measured by infectious center in situ plaque hybridization in monkey BSC-1 cells. The sequences required for the enhancement of recombination by linear DNA reside within the SV40 replication origin/regulatory region (nucleotides 5,171 to 5,243/0 to 128). Linearization of phi X174 RFI DNA did not increase the recombination frequency. The SV40/phi X174 recombinant structures arising from transfections supplemented with linear forms of origin-containing SV40 DNA contained phi X174 DNA sequences interspersed within tandem head-to-tail repeats derived from the recombination-enhancing linear DNA. Evidence is presented that the tandem repeats are not formed by homologous recombination and that linear forms of SV40 DNA must compete with circular SV40 DNA for the available T antigen to enhance recombination. We propose that the enhancement of recombination by linear SV40 DNA results from the entry of that DNA into a rolling circle type of replication pathway which generates highly recombinogenic intermediates.

scholarly journals Identification of Simian virus 40 promoter DNA sequences capable of conferring restriction endonuclease hypersensitivity.

1996 ◽  
Vol 70 (6) ◽  
pp. 3416-3422 ◽  
Author(s):  
R Hermansen ◽  
M A Sierra ◽  
J Johnson ◽  
M Friez ◽  
B Milavetz
Keyword(s):  
Restriction Endonuclease ◽  
Dna Sequences ◽  
Simian Virus 40 ◽  
Simian Virus ◽  
Promoter Dna

scholarly journals Chromatin Structure of the Simian Virus 40 Late Promoter: a Deletional Analysis

1999 ◽  
Vol 73 (3) ◽  
pp. 1990-1997 ◽  
Author(s):  
Michael Friez ◽  
RaeJean Hermansen ◽  
Barry Milavetz
Keyword(s):  
Dna Sequences ◽  
Simian Virus 40 ◽  
Simian Virus ◽  
Late Promoter ◽  
Reporter System ◽  
Free Region ◽  
Restriction Sites ◽  
Unique Restriction ◽  
Minimal Sequence ◽  
Nuclease Sensitivity

ABSTRACT The goal of this study was to determine the minimal sequence within the simian virus 40 (SV40) late promoter region, nucleotides (nt) 255 to 424, capable of phasing nucleosomes as measured by its ability to confer the greatest endonuclease sensitivity on adjacent DNA sequences. To identify the minimal sequence, a deletional analysis of the late region was performed by utilizing a SV40 recombinant reporter system. The reporter system consisted of a series of unique restriction sites introduced into SV40 at nt 2666. The unique restriction sites allowed the insertion of test sequences as well as measurement of conferred endonuclease sensitivity. The results of the deletional analysis demonstrated that constructs capable of conferring the greatest nuclease sensitivities consistently included nt 255 to 280. The activator protein 4 (AP-4) and GTIIC transcription factor binding sequences lie within this region and were analyzed individually. Their abilities to confer nuclease sensitivity upon the reporter nearly matched that of the entire late domain. These results suggest that transcription factors AP-4 and transcription-enhancing factor which binds the GTIIC sequence are able to confer significant levels of nuclease sensitivity and are likely involved in the formation of the SV40 nucleosome-free region.

Molecular analysis of enhanced replication of UV-damaged simian virus 40 DNA in UV-treated mammalian cells

1988 ◽  
Vol 8 (6) ◽  
pp. 2428-2434
Author(s):  
J M Treger ◽  
J Hauser ◽  
K Dixon
Keyword(s):  
Uv Radiation ◽  
Uv Irradiation ◽  
Simian Virus 40 ◽  
Simian Virus ◽  
Viral Dna ◽  
Repair Capacity ◽  
Pyrimidine Dimers ◽  
Infected Cells ◽  
Replicative Intermediates ◽  
Kinetics Of

Irradiation of simian virus 40 (SV40)-infected cells with low fluences of UV light (20 to 60 J/m2, inducing one to three pyrimidine dimers per SV40 genome) causes a dramatic inhibition of viral DNA replication. However, treatment of cells with UV radiation (20 J/m2) before infection with SV40 virus enhances the replication of UV-damaged viral DNA. To investigate the mechanism of this enhancement of replication, we analyzed the kinetics of synthesis and interconversion of viral replicative intermediates synthesized after UV irradiation of SV40-infected cells that had been pretreated with UV radiation. This enhancement did not appear to be due to an expansion of the size of the pool of replicative intermediates after irradiation of pretreated infected cells; the kinetics of incorporation of labeled thymidine into replicative intermediates were very similar after irradiation of infected control and pretreated cells. The major products of replication of SV40 DNA after UV irradiation at the low UV fluences used here were form II molecules with single-stranded gaps (relaxed circular intermediates). There did not appear to be a change in the proportion of these molecules synthesized when cells were pretreated with UV radiation. Thus, it is unlikely that a substantial amount of DNA synthesis occurs past pyrimidine dimers without leaving gaps. This conclusion is supported by the observation that the proportion of newly synthesized SV40 form I molecules that contain pyrimidine dimers was not increased in pretreated cells. Pulse-chase experiments suggested that there is a more efficient conversion of replicative intermediates into form I molecules in pretreated cells. This could be due to more efficient gap filling in relaxed circular intermediate molecules or to the release of blocked replication forks. Alternatively, the enhanced replication observed here may be due to an increase in the excision repair capacity of the pretreated cells.

Bovine papilloma virus contains an activator of gene expression at the distal end of the early transcription unit

1983 ◽  
Vol 3 (6) ◽  
pp. 1108-1122
Author(s):  
M Lusky ◽  
L Berg ◽  
H Weiher ◽  
M Botchan
Keyword(s):  
Thymidine Kinase ◽  
Dna Sequences ◽  
Base Pair ◽  
Simian Virus 40 ◽  
Simian Virus ◽  
Stable Transformation ◽  
High Molecular Weight Dna ◽  
Papilloma Virus ◽  
Recombinant Plasmids ◽  
Bovine Papilloma Virus

Bovine papilloma virus (BPV) contains a cis-acting DNA element which can enhance transcription of distal promoters. Utilizing both direct and indirect transient transfection assays, we showed that a 59-base-pair DNA sequence from the BPV genome could activate the simian virus 40 promoter from distances exceeding 2.5 kilobases and in an orientation-independent manner. In contrast to the promoter 5'-proximal localization of other known viral activators, this element was located immediately 3' to the early polyadenylation signal in the BPV genome. Deletion of these sequences from the BPV genome inactivated the transforming ability of BPV recombinant plasmids. Orientation-independent reinsertion of this 59-base-pair sequence, or alternatively of activator DNA sequences from simian virus 40 or polyoma virus, restored the transforming activity of the BPV recombinant plasmids. Furthermore, the stable transformation frequency of the herpes simplex virus type 1 thymidine kinase gene was enhanced when linked to restriction fragments of BPV DNA which included the defined activator element. This enhancement was orientation independent with respect to the thymidine kinase promoter. The enhancement also appeared to be unrelated to the establishment of the recombinant plasmids as episomes, since in transformed cells these sequences are found linked to high-molecular-weight DNA. We propose that the enhancement of stable transformation frequencies and the activation of transcription units are in this case alternate manifestations of the same biochemical events.

Simian virus 40 major late promoter: an upstream DNA sequence required for efficient in vitro transcription

1984 ◽  
Vol 4 (1) ◽  
pp. 133-141
Author(s):  
J Brady ◽  
M Radonovich ◽  
M Thoren ◽  
G Das ◽  
N P Salzman
Keyword(s):  
Dna Sequence ◽  
Dna Sequences ◽  
Simian Virus 40 ◽  
Promoter Sequence ◽  
In Vitro Transcription ◽  
Simian Virus ◽  
Upstream Promoter ◽  
Upstream Promoter Sequence

We have previously identified an 11-base DNA sequence, 5'-G-G-T-A-C-C-T-A-A-C-C-3' (simian virus 40 [SV40] map position 294 to 304), which is important in the control of SV40 late RNA expression in vitro and in vivo (Brady et al., Cell 31:625-633, 1982). We report here the identification of another domain of the SV40 late promoter. A series of mutants with deletions extending from SV40 map position 0 to 300 was prepared by nuclease BAL 31 treatment. The cloned templates were then analyzed for efficiency and accuracy of late SV40 RNA expression in the Manley in vitro transcription system. Our studies showed that, in addition to the promoter domain near map position 300, there are essential DNA sequences between nucleotide positions 74 and 95 that are required for efficient expression of late SV40 RNA. Included in this SV40 DNA sequence were two of the six GGGCGG SV40 repeat sequences and an 11-nucleotide segment which showed strong homology with the upstream sequences required for the efficient in vitro and in vivo expression of the histone H2A gene. This upstream promoter sequence supported transcription with the same efficiency even when it was moved 72 nucleotides closer to the major late cap site. In vitro promoter competition analysis demonstrated that the upstream promoter sequence, independent of the 294 to 304 promoter element, is capable of binding polymerase-transcription factors required for SV40 late gene transcription. Finally, we show that DNA sequences which control the specificity of RNA initiation at nucleotide 325 lie downstream of map position 294.

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