Comparative Studies of Wild-type and Cold-mutant (Temperature-sensitive) Influenza Viruses: Independent Segregation of Temperature-sensitivity of Virus Replication from Temperature-sensitivity of Virion Transcriptase Activity during Recombination of Mutant A/Ann Arbor/6/60 with Wild-type H3N2 Strains

Kendal, Alan P.; Cox, Nancy J.; Galphin, Judith C.; Maassab, Hunein F.

doi:10.1099/0022-1317-44-2-443

Cited by 22 publications

(14 citation statements)

References 13 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…However, as it has been demonstrated that RNA segment 1 of fowl plague virus Rostock is functionally equivalent to RNA segment 2 of the WSN strain of influenza virus (Scholtissek, 1978(Scholtissek, , 1979 there is a clear correlation between the results presented here and those obtained in previous studies with influenza WSN (Mowshowitz & Ueda, 1976: Mowshowitz, 1978, influenza A/Ann Arbor/6/60 (Kendal et al, 1978(Kendal et al, , 1979 and fowl plague virus Weybridge (Ghendon et al, 1975). In all cases, a protein encoded in the genome RNA segment functionally equivalent to influenza WSN RNA segment 2 is essential for the function of the virion transcriptase.…”

supporting

confidence: 74%

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

View full text Add to dashboard Cite

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...

show abstract

supporting

confidence: 74%

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

View full text Add to dashboard Cite

show abstract

“…However, it is also possible that events necessary for mRNAprimed transcription (such as endonuclease activity or associated conformational changes in the polymerase) are not completely decoupled from transcription during ApG-primed synthesis. Consistent with the latter possibility, several ts virus mutants have been found with lesions in the PB2 gene that affect dinucleotide-primed transcription (Kendal et al, 1979;Nichol et al, 1981). Nevertheless, the primary effect of the F5 serum on normally primed synthesis seems to be inhibition of the endonuclease.…”

Section: Discussionsupporting

confidence: 51%

Inhibition of the influenza virus RNA-dependent RNA polymerase by antisera directed against the carboxy-terminal region of the PB2 subunit

Blok

Cianci

Tibbles

et al. 1996

Journal of General Virology

View full text Add to dashboard Cite

The influenza virus RNA polymerase consists of a heterotrimeric complex of the PB1, PB2 and PA proteins, with the PB2 subunit responsible for recognizing 5' cap structures on the host cell RNAs used as primers for virus mRNA synthesis. To investigate further the role PB2 plays in mRNA synthesis, a set ofpolyclonal antisera raised against defined regions of the protein were tested for their ability to inhibit the virion transcriptase. All five sera were of sufficient titre to immunoprecipitate PB2 and four were capable of recognizing polymerase complexes containing PB 1 and PA. However, only the serum raised against the carboxy terminus of PB2 (F5) substantially inhibited polymerase activity. This serum drastically reduced synthesis primed by globin mRNA, but only partially inhibited transcription primed by the dinucleotide ApG, or ApG and cap analogue. The preferential inhibition of globinprimed synthesis did not result from interference with cap recognition, as serum F5 did not reduce labelling of PB2 in a photoaffinity cap-binding assay. However, IgG and Fab fragments from F5 were found to inhibit virion endonuclease activity. This suggests that the C terminus of PB2 plays a crucial role in transcription initiation and implicates PB2 in endonuclease activity.

show abstract

“…At 25 ° C the activities of any influenza wt strains do not surpass their activities at the optimal temperature (11,22). The ca A/AA/6/60 variant and any ca reassortants containing the A/AA/6/60 PB 2 gene, however, exhibited identical activities at both 25 ° and 33 ° C in our assay.…”

Section: Discussionmentioning

confidence: 64%

Biological characteristics of a cold-adapted influenza A virus mutation residing on a polymerase gene

Odagiri

Tosaka

Ishida

et al. 1986

Archives of Virology

Self Cite

View full text Add to dashboard Cite

The biological function of a cold-adapted (ca) mutation residing on the PB2 gene of an influenza A/Ann Arbor/6/60 (A/AA/6/60) ca variant virus in the viral replication cycle at 25 degrees C was studied. The viral polypeptide synthesis of A/AA/6/60 ca variant at 25 degrees C was evident approximately 6 hours earlier than the wild type (wt) virus and yielded twice as many products. The quantitative analysis of viral complementary RNA (cRNA), synthesized in the presence of cycloheximide, revealed that A/AA/6/60 ca variant and a single gene reassortant that contains only the PB2 gene of the ca variant with remaining genes of the wt virus produced equal amount of cRNA at 25 degrees and 33 degrees C, which was an amount approximately four fold greater than the wt virus' cRNA synthesized at 25 degrees C. These results strongly suggest that the ca mutation residing on the PB2 gene of A/AA/6/60 ca variant affects the messenger RNA synthesis at 25 degrees C in the primary transcription.

show abstract

Cited by 22 publications

References 13 publications

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

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

Inhibition of the influenza virus RNA-dependent RNA polymerase by antisera directed against the carboxy-terminal region of the PB2 subunit

Biological characteristics of a cold-adapted influenza A virus mutation residing on a polymerase gene

Contact Info

Product

Resources

About