The effect of Mengovirus infection on the activity of the DNA-dependent RNA polymerase of L-cells. II. Preliminary data on the inhibitory factor

Balandin, I. G.; Franklin, Richard M.

doi:10.1016/0006-291x(64)90097-x

Cited by 32 publications

(13 citation statements)

References 6 publications

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“…Taken together, these data indicate that the virus interacts with cellular regulatory mechanisms of RNA synthesis, not with tran scription per se. This idea is consistent with the observation that different cell lines respond differently in their RNA synthesis to the in hibitory activity of the same picornavirus [6], Earlier work has shown that viral protein synthesis is necessary to induce the transcrip tional inhibition [7,8], Others have failed to demonstrate a direct inhibitory activity of viral proteins in in vitro transcription sys tems [9, 10a]. Recently, Bossart et al [10b] were able to show in vitro that cytoplasmic extracts (S30) of poliovirus-infected cells in hibit RNA synthesis in isolated nuclei.…”

supporting

confidence: 75%

“…It has been well known since the early 1960s that picornaviruses inducean inhibition of host cell transcription [3,21,22] and that virusspecific protein synthesis appears to be respon sible for RNA synthesis shut-off [8]. Since it has been found that cytoplasmic extracts of poliovirus-infected cells can inhibit transcrip tion in uninfected, isolated nuclei [10b], the next step was to see if poliovirus proteins could be demonstrated within the host cell nucleus.…”

Section: Discussionmentioning

confidence: 99%

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Accumulation of Poliovirus Proteins in the Host Cell Nucleus

et al. 1982

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Poliovirus (type 1, Mahoney) proteins were transported into the nuclei of HEp-2 cells, as demonstrated by means of electron microscopic autoradiography. Quantitative determinations of the proteins by electrophoresis showed that the relative amounts of precursor proteins (la, 3b, lb), of some of the intermediate polypeptides (3a, 3c, 5b, 2), and of one end product (VP1) were higher in the nucleus than in the cytoplasm. Other proteins (VPO, YP2, X, 4, 6a) were found in the same relative amounts in the nucleus as in the cytoplasm, whereas two proteins (VP3 and 6b) were excluded from the nucleus. The findings are discussed in view of the recently demonstrated inhibitory activity exerted by cytoplasmic extracts of poliovirus-infected cells on transcription in isolated nuclei of uninfected cells.

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supporting

confidence: 75%

Section: Discussionmentioning

confidence: 99%

Accumulation of Poliovirus Proteins in the Host Cell Nucleus

et al. 1982

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“…These and results from in vitro experiments [7] pres ent evidence that poliovirus interacts with host RNA transcription initiation. Host RNA synthesis inhibition by picornaviruses is induced by viral proteins [8]; the inhibiting protein(s), however, have never been iden tified. Specific poliovirus proteins were shown to accumulate in the host cell nuclei [9,10],…”

mentioning

confidence: 99%

Accumulation of Poliovirus Proteins in Uninfected Isolated HEp-2 Cell Nuclei in vitro

Bossart¹,

Egger²,

Rasser³

et al. 1984

Intervirology

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Poliovirus (type 1, Mahoney) proteins preferentially inhibited in vitro RNA polymerase II activity in uninfected isolated HEp-2 cell nuclei, as demonstrated by electron microscopic autoradiography. Structural integrity of the nuclei and preincubation of nuclei and poliovirus proteins in vitro prior to addition of [³H]-UTP were prerequisites for differential inhibition. During in vitro incubation, [³H]-labeled poliovirus proteins were shown to accumulate in the uninfected isolated nuclei. Similar poliovirus proteins accumulated in the nuclei to higher relative amounts as compared to the outside or were excluded from the nuclei after in vitro incubation, as in intact poliovirus-infected cells. Since the in vitro transcription system of isolated nuclei exhibited two characteristics of poliovirus infection, i.e., preferential inhibition of RNA polymerase II activity and accumulation of poliovirus proteins in the nuclei, it may be useful to study host RNA synthesis inhibition.

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“…Balandin & Franklin (1964) postulated that in mengovirus-infected L cells, a virus protein synthesized in the cytoplasm migrates into the host cell nucleus and interacts with host nuclear functions. In fact, we have demonstrated an accumulation of poliovirus proteins in the host cell nucleus (Bienz et al, 1982).…”

Section: Discussionmentioning

confidence: 99%

“…Lipid or lipoprotein (Ho & Washington, 1971) as well as virus proteins (Balandin & Franklin, 1964) have been postulated to be the inhibiting agent(s). We found an accumulation of poliovirus proteins in the host cell nucleus as judged by PAGE and by EM autoradiography (Bienz et al, 1982).…”

Section: Introductionmentioning

confidence: 99%

Poliovirus-induced Inhibition of Host RNA Synthesis Studied in Isolated HEp-2 Cell Nuclei

Bossart

Egger

Rasser

et al. 1982

Journal of General Virology

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SUMMARYNuclei isolated from uninfected HEp-2 cells synthesized RNA for 60 to 90 min. The individual RNA polymerase activities were determined by u-amanitin differential inhibition and the RNA products characterized by electron microscope (EM) autoradiography and sucrose gradient centrifugation. In nuclei prepared from poliovirusinfected cells, the capacity to synthesize RNA in vitro decreased with time after infection. RNA polymerase II activity (hnRNA synthesis) was preferentially inhibited more than was the polymerase I activity (rRNA synthesis). Poliovirus-infected cytoplasm (S-30) inhibited in vitro RNA synthesis in uninfected nuclei by selectively affecting the polymerase II activity. Selective inhibition of hnRNA synthesis by the crude extracts could be monitored by EM autoradiography directly. Determinations of individual RNA polymerase activities by differential ~-amanitin inhibition were done only after treatment of the infected cytoplasm with micrococcal nuclease to abolish virus RNA replication. Selective inhibition of hnRNA synthesis depended on preincubation of the nuclei together with the infected cytoplasm, indicating that inhibitory substances from the infected cytoplasm entered the nuclei. Isolated nuclei therefore provide a useful system for studying the nature of the inhibitor(s) and of host RNA synthesis inhibition by picornaviruses.

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The effect of Mengovirus infection on the activity of the DNA-dependent RNA polymerase of L-cells. II. Preliminary data on the inhibitory factor

Cited by 32 publications

References 6 publications

Accumulation of Poliovirus Proteins in the Host Cell Nucleus

Accumulation of Poliovirus Proteins in the Host Cell Nucleus

Accumulation of Poliovirus Proteins in Uninfected Isolated HEp-2 Cell Nuclei in vitro

Poliovirus-induced Inhibition of Host RNA Synthesis Studied in Isolated HEp-2 Cell Nuclei

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