The control of SV40 transcription during a lytic infection: late RNA synthesis in the presence of inhibitors of DNA replication

Rosenthal, Leonard J.; Brown, M.

doi:10.1093/nar/4.3.551

Cited by 23 publications

(11 citation statements)

References 26 publications

(31 reference statements)

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“…3 and unpublished results obtained by immunoprecipitation) which could not be detected on stained gels. Synthesis of very small amounts of late viral mRNAs coding for SV40 or polyoma virus capsid proteins, in the presence of araC or FdUrd, has been reported (1,13). In contrast, no evidence for synthesis of viral capsid proteins could be detected in secondary monkey kidney cultures infected with SV40 in the presence of araC.…”

Section: Methodsmentioning

confidence: 99%

Simian virus 40 and polyoma virus stimulate overall cellular RNA and protein synthesis.

Khandjian¹,

Matter²,

Leonard³

et al. 1980

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

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In lytic infection with simian virus 40 and polyoma virus of monkey and mouse cells in tissue culture, synthesis of the viral tumor (T) antigens (T antigens) is rapidly followed by a mitogenic response of the host cell. The latter begins with virus-induced stimulation of overall cellular RNA and protein synthesis, leading to a substantial increase in cytoplasmic and nuclear RNA and protein. Stimulation begins within 1 hr after onset of T-antigen synthesis and also occurs if virus-induced DNA synthesis is blocked by metabolic inhibitors. The broad spectrum of biological and molecular effects induced by simian virus 40 and polyoma virus is, at least phenotypically, reminiscent of the pleiotropic impact exerted on target cells by nonviral mitogens and by certain growth-promoting steroid and polypeptide hormones.Simian virus 40 (SV40) and polyoma virus induce a lytic infection in permissive cells and an abortive ("transforming") infection in nonpermissive cells. These infections exhibit considerable similarity (for details and references, see ref. 1). Expression of the early viral gene-i.e., synthesis of virus-specific early 19S mRNAs and of the tumor antigens (T antigens), is rapidly followed by a mitogenic reaction of the host cell. This reaction includes virus-induced stimulation of overall cellular RNA synthesis and an increase in total, mainly ribosomal RNA, activation of the cellular DNA-synthesizing apparatus and duplication of the host cell chromatin (S phase). In nonpermissive cells, virus-induced S phase is followed by prophase and mitosis but no viral DNA is replicated. In permissive cells, S phase is paralleled by replication of viral DNA as a nucleohistone and by production of progeny virus and is followed by cell death (lysis). The early events of infection, including the activation of the cellular DNA-synthesizing apparatus, also occur if virus-induced DNA synthesis is blocked by metabolic in-In this paper we report that, in lytic infection, SV40-and polyoma-induced stimulation of cellular RNA synthesis is paralleled by stimulation of cellular protein synthesis which also occurs in the presence of araC or FdUrd.MATERIALS AND METHODS Primary mouse kidney (2), secondary monkey kidney, and CV-1 (a monkey kidney cell line) cultures were grown in 10-cm-diameter plastic dishes in reinforced Eagle's medium ("culture medium") containing 10% fetal bovine serum (GIBCO) (3,4 To separate cells into cytoplasmic and nuclear fractions, cultures were incubated for 10 min at 40C in lysis buffer (300 mM sucrose/10 mM Tris-HCI, pH 7.4/5 mM NaCI/3 mM MgCl2/0.5% Nonidet P-40), 1 ml per dish. The cells were scraped from the plates and passed through a syringe (20-gauge needle) seven times, and the lysate was centrifuged at 3000 X g for 15 min at 40C.To extract proteins, unfractionated cultures were suspended, at 1.0 ml per dish, in 1% NaDodSO4/1 mM NaH2PO4, pH 8.5; nuclear pellets were suspended in 0.25 ml per dish. To cytoplasmic fractions (1 ml per dish) 50 ,ul of 20% (wt/vol) NaDodSO4 was added. The lysates were...

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Section: Methodsmentioning

confidence: 99%

Simian virus 40 and polyoma virus stimulate overall cellular RNA and protein synthesis.

Khandjian¹,

Matter²,

Leonard³

et al. 1980

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

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“…The SV40 early protein large T antigen is largely responsible for facilitating the replication of the viral genome and directing the synthesis of the capsid proteins VP1, VP2, and VP3 (3,11,16,26,29,32). Upon synthesis, VP1 readily forms pentamers that contain a single copy of either VP2 or VP3 within their central cavity (1,9).…”

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confidence: 99%

Simian Virus 40 Late Proteins Possess Lytic Properties That Render Them Capable of Permeabilizing Cellular Membranes

Daniels¹,

Rusan²,

Wilbuer³

et al. 2006

J Virol

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Many nonenveloped viruses have evolved an infectious cycle that culminates in the lysis or permeabilization of the host to enable viral release. How these viruses initiate the lytic event is largely unknown. Here, we demonstrated that the simian virus 40 progeny accumulated at the nuclear envelope prior to the permeabilization of the nuclear, endoplasmic reticulum, and plasma membranes at a time which corresponded with the release of the progeny. The permeabilization of these cellular membranes temporally correlated with late protein expression and was not observed upon the inhibition of their synthesis. To address whether one or more of the late proteins possessed an inherent capacity to induce membrane permeabilization, we examined the permeability of Escherichia coli that separately expressed the late proteins. VP2 and VP3, but not VP1, caused the permeabilization of bacterial membranes. Additionally, VP3 expression resulted in bacterial cell lysis. These findings demonstrate that VP3 possesses an inherent lytic property that is independent of eukaryotic signaling or cell death pathways.

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“…Earlier studies (26) showed, by cesium chloride/propidium diiodide equilibrium centrifugation and by alkaline sucrose sedimentation, that essentially no tsA58 DNA replication occurs at >40°. A more sensitive method of detection of DNA replication is DNA-DNA filter hybridization of pulse-labeled DNA, in which as few as 200-250 virus-specific input counts could be detected with an efficiency of about 10% (26).…”

mentioning

confidence: 99%

Characterization of early simian virus 40 transcriptional complexes: late transcription in the absence of detectable DNA replication.

Ferdinand¹,

Brown²,

Khoury³

1977

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

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Isolation of early viral transcriptional complexes and incorporation in vitro of radiolabeled precursors into nascent RNA has permitted an analysis of early simian virus 40 (SV40) transcription. Under conditions such that viral DNA replication was undetectable, both early and late SV40 RNA were synthesized. This finding provides evidence that viral DNA replication is not an absolute requirement for late transcription and supports earlier observations that late viral RNA is synthesized in SV40-infected nonpermissive mouse cells. The majority of the early viral transcriptional activity can be solubilized, indicating that a substantial portion of this RNA is transcribed from free rather than integrated templates. Sedimentation analysis of the transcriptional complexes resulted in the detection of two separate peaks of activity, suggesting the possibility of two distinct types of early SV40 templates.It has been known for some time that simian virus 40 (SV40) RNA is synthesized in two phases (1-3). Prior to viral DNA replication, most of the viral mRNA appears to be transcribed from the early DNA strand (4-6). Subsequently, the bulk of viral transcription occurs from the late DNA strand and is processed into late 16S and 19S viral mRNAs (7,8). It was generally assumed that this late phase of viral transcription was inextricably linked to viral DNA replication (see refs. 9 and 10). The absence of the late SV40 transcripts in most SV40-transformed cell lines supported this concept (11). Nevertheless, we consistently found late viral RNA sequences in abortively infected mouse cells, in which SV40 DNA replication could not be detected (5, 12). Furthermore, recent studies with early temperature-sensitive mutants of SV40 grown at 410 (the nonpermissive temperature) indicated the presence of a small but reproducible fraction of late virus-specific cytoplasmic RNA (13). This led us to speculate that viral DNA replication was not an absolute requirement for the transcription of sequences on the late SV40 DNA strand. It was difficult to test this hypothesis directly, because of the low levels of late virus-specific RNA found prior to viral DNA replication (1-6, 14) and the potential rapidity and efficiency of the RNA processing system (15).We recently described the adaptation of an in vitro transcriptional system (16) established by Gariglio and Mousset (17). The supernatant fraction of SV40-infected cell nuclei, lysed in the presence of Sarkosyl, actively incorporates radiolabeled ribonucleotide triphosphates, and the newly synthesized RNA can be easily analyzed. Although it is unclear whether putative termination signals are recognized under such incorporation conditions, the absence of reinitiation allows one to assess accurately the frequency of in vivo transcription of the two viral DNA strands (16). Further advantages of this in vitro transcriptional system include: (i) the enrichment of virus-specific RNA sequences in the nuclear supernatant fraction; (ii) an increase in the specific activity of the viral RNA t...

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The control of SV40 transcription during a lytic infection: late RNA synthesis in the presence of inhibitors of DNA replication

Cited by 23 publications

References 26 publications

Simian virus 40 and polyoma virus stimulate overall cellular RNA and protein synthesis.

Simian virus 40 and polyoma virus stimulate overall cellular RNA and protein synthesis.

Simian Virus 40 Late Proteins Possess Lytic Properties That Render Them Capable of Permeabilizing Cellular Membranes

Characterization of early simian virus 40 transcriptional complexes: late transcription in the absence of detectable DNA replication.

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