Subclasses of ribonucleoproteins in influenza virus-infected cells

Caliguiri, Lawrence A.; Gerstein, Hal

doi:10.1016/0042-6822(78)90339-2

Cited by 16 publications

(15 citation statements)

References 37 publications

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“…However, since NP forms a heterogeneous collection of structures [free protein, structural RNP (RNA-), replicative RNP (RNA+); Caliguiri & Gerstein, 1978;P. Rees & N. J. Dimmock, unpublished results], significant changes in phosphorylation in one of these species may be concealed and cannot yet be ruled out; neither do we have information about the intramolecular distribution of phosphate groups in nuclear and cytoplasmic NPs.…”

Section: -2mentioning

confidence: 99%

“…The influenza virus-infected cell is a well-characterized system which lends itself to the study of the function of virus phosphoproteins, particularly since the phosphoprotein NP is involved in a number of activities. For instance, it forms replicative and structural nucleoprotein complexes (Caliguiri & Gerstein, 1978), interacts with both virion and complementary RNA (Pons, 1975), and is transported into the nucleus (Breitenfeld & Sch~ifer, 1957;Taylor et al, 1969Taylor et al, , 1970Lazarowitz et al, 1971;Krug & Etkind, 1973;Hay & Skehel, 1975) and subsequently back into the cytoplasm (Flawith & Dimmock, 1979). We have investigated the phosphorylation of influenza proteins in vivo to determine whether phosphorylation might affect the movement of NP between the nucleus and cytoPlasm.…”

Section: Phosphorylation Of Influenza Virus Nueleoprotein In Vivomentioning

confidence: 99%

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Phosphorylation of Influenza Virus Nucleoprotein in vivo

Petri

Dimmock

1981

Journal of General Virology

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Section: -2mentioning

confidence: 99%

Section: Phosphorylation Of Influenza Virus Nueleoprotein In Vivomentioning

confidence: 99%

Phosphorylation of Influenza Virus Nucleoprotein in vivo

Petri

Dimmock

1981

Journal of General Virology

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“…Several groups of investigators have attempted to identify the virus-specific proteins involved in the transcription of influenza virus RNA. Both fractionation studies of virus ribonucleoprotein complexes (Caliguiri & Compans, 1974: Caliguiri & Gerstein, 1978: Inglis et al, 1976 and the study of RNA synthesis in cells infected with influenza virus ts mutants (Sugiura et al, 1975: Krug et al,, 1975: Scholtissek et al, 1974: Scholtissek & Bowles, 1975: Scholtissek, t978, 1979: Ghendon et aL, 1973: Barry & Mahy, 1979 have indicated a requirement for the P proteins and the nucleocapsid protein in transcription of influenza virus RNA, although the precise function of these proteins remained unclear.…”

mentioning

confidence: 99%

Evidence for the Involvement of Influenza A (Fowl Plague Rostock) Virus Protein P2 in ApG and mRNA Primed in vitro RNA Synthesis

Nichol

Penn

Mahy

1981

Journal of General Virology

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SUMMARYEleven temperature-sensitive (ts) mutants of influenza A (fowl plague, Rostock) virus were analysed for in vitro RNA transcriptase activity in reactions primed by ApG or globin rnRNA at 31 °C or at 40.5 °C, the restrictive temperature for ts mutant growth. Only those ts mutants studied which were defective in RNA segment 1, coding for the virion P2 protein, were defective in RNA transcriptase activity when compared to wild-type virus. Mutants having a defect in the P2 protein had no significant RNA transcriptase activity in reactions at 40.5 °C primed by globin mRNA. However, one mutant showed RNA transcriptase activity similar to wild-type virus at 40.5 °C when ApG (0.3 raM) was used as primer. The results suggest that influenza (fowl plague, Rostock) P2 protein is directly involved in the mRNA priming reaction, as well as in the RNA transcription reaction in vitro.The eight distinct negative-strand RNA segments of fowl plague virus, an avian influenza A virus, can be transcribed in vitro by the virion-bound RNA-dependent RNA polymerase (for reviews, see Barry & Mahy, 1979~ Hay & Skehel, 1979, Under optimal conditions the products of the influenza virion RNA-dependent RNA polymerase reaction have many of the properties of virus mRNA synthesized in infected cells, in that they are approx, full-length, polyadenylated transcripts of each genome segment (Plotch & Krug, 1977, 1978: Hay et al., 1977. However, the structures of in vivo and in vitro synthesized mRNA do differ in that the in vitro mRNAs are shorter and lack a 5' methylated cap structure (Hay et al., 1977. Furthermore, primary transcription in vivo is sensitive to the drugs actinomycin D and alpha-amanitin (inhibitors of host cell DNA-dependent RNA polymerase ll), whereas transcription in vitro is insensitive to these drugs (Scholtissek & Rott, 1970;Penhoet et aL, 1971;Mahy et aL, 1972;Lamb & Choppin, 1977;Spooner & Barry, 1977). In addition, the influenza virion transcriptase was shown to possess another unusual property in that a number of guanosine related compounds, particularly the dinucleoside adenyl (3' to 5') guanosine (ApG), stimulate RNA synthesis in vitro by acting as primers for RNA synthesis (McGeoch & Kitron, 1975;Plotch & Krug, 1977).Recently, an explanation for these findings became apparent when it was reported that various eukaryotic mRNAs stimulate the in vitro transcription reaction (Bouloy et al., 1978) and that the 5'-terminal cap structure plus 9 to 15 nucleotides is transferred to the 5' end of the transcript in vitro (Plotch et al., 1979;Bouloy et al., 1980. The authors suggested that capped host cell RNA molecules are used as 'primers' for synthesis of influenza mRNAs in vivo and that the continued synthesis of these primer molecules is the alpha-amanitin-sensitive step in influenza virus replication (Kruget aL, 1979). In an attempt to determine which virion proteins participate in this priming reaction, we have studied the ApG and globin mRNA-primed in vitro transcriptase activities of a number of genetically characteriz...

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“…In considering the movements of NP to the nucleus, there is the added complication that some of it is associated with viral RNA to form RNP complexes in both the virus particle (Pons et al, 1969;Duesberg, 1969;Compans et al, 1972;Rees & Dimmock, 1981b) and in infected cells (Pons, 1971(Pons, , 1975Caliguiri & Gerstein, 1978;. These RNPs are found in both the cytoplasm and the nucleus of infected cells (Krug, 1971) and appear able to cross the nuclear envelope in either direction (Flawith & Dimmock, 1979;Hudson et al, 1978).…”

Section: Discussionmentioning

confidence: 99%

“…Analysis of radiolabelled proteins following the fractionation of infected cells showed that after synthesis in the cytoplasm, the majority of NP migrates to the nucleus where it associates with the nucleoplasm (Taylor et al, 1969(Taylor et al, , 1970Lazarowitz et al, 1971;Krug & Etkind, 1973;Hay & Skehel, 1975;Flawith & Dimmock, 1979;Briedis et al, 1981). The situation is complicated because NP is found in both nucleus and cytoplasm as 'free' protein and associated with viral RNA, and the latter ribonucleoproteins (RNPs) contain both negative and positive strand viral RNA (Pons, 1971(Pons, , 1975Caliguiri & Gerstein, 1978;. Also NP appears to be able to cross the nuclear envelope in either direction (Hudson et al, 1978;Flawith & Dimmock, 1979).…”

Section: Introductionmentioning

confidence: 99%

Location of Influenza Virus M, NP and NS1 Proteins in Microinjected Cells

Davey

Colman

Dimmock

1985

Journal of General Virology

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SUMMARYWhen microinjected as cloned DNA, the nucleoprotein (NP) of influenza virus A/NT/60/68 (H3N2) accumulated in the nuclei of Xenopus laevis oocytes, and cultured cells of rodent and primate origin. This accumulation appeared to be specific and a property of the NP itself (or conceivably NP in association with unknown cellular constituents) since no other influenza virus components were present in DNA-injected cells. In the oocyte nucleus, clonally derived NP achieved an eightfold concentration over that in the cytoplasm. Such NP was full-length as judged by its mobility during PAGE and had the native conformation of H3N2 virus NP according to its reaction with a panel of monoclonal antibodies. NP appeared to be in the soluble fraction of the nucleus as it did not sediment under conditions which removed particulate matter from nuclear extracts. Microinjection of extracts of chick embryo fibroblast cells infected with A/FPV/Rostock/34 (H7N1) showed that exogenous NP had an affinity for the nucleus similar to that synthesized intracellularly from cloned NP DNA. This conclusion was supported by an experiment in which cloned NP from the oocyte nucleus re-entered the nucleus after injection into the cytoplasm of fresh oocytes. Injection of mRNA, extracted from chick embryo fibroblast cells infected with A/FPV/Rostock/34, into oocytes directed the synthesis of the viral proteins M (Mr 28000), and NS1 (MT 27000) as well as NP (Mr 56000). While NP from this source concentrated in the nucleus as before, M merely associated with the nucleus without exceeding the cytoplasmic level. Even more remarkable was NS1 ; although in injected cells this protein is concentrated in nucleoli, in microinjected oocytes its nuclear concentration was threefold less than that in the cytoplasm, despite the very large number (> 1500) of nucleoli present in Xenopus oocytes. It seems likely that the karyophilic nature of M and NS1, unlike that of NP, is a property not of the proteins themselves, but of a complex which they form with some other product of the infected cell. These findings were repeated when extracts from infected chick embryo cells containing NP, M and NS1 proteins radiolabelled in vivo, were injected into the cytoplasm of oocytes.

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Subclasses of ribonucleoproteins in influenza virus-infected cells

Cited by 16 publications

References 37 publications

Phosphorylation of Influenza Virus Nucleoprotein in vivo

Phosphorylation of Influenza Virus Nucleoprotein in vivo

Evidence for the Involvement of Influenza A (Fowl Plague Rostock) Virus Protein P2 in ApG and mRNA Primed in vitro RNA Synthesis

Location of Influenza Virus M, NP and NS1 Proteins in Microinjected Cells

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