Search PubMed⌕ Search

SEARCH · Search PubMed

Results for “Polyproteins”

Search indexed PubMed citations on genomics, clinical trials, systematic reviews and public health. Explore titles, authors and supplied subject terms, then open the PubMed record.

Quote a phrase for an exact phrase match. Source license links do not imply unrestricted reuse.

At least 487 records · Page 27Linked to original sources
PubMed1984

Impaired cleavage of the joint gag-pol polyprotein precursor and virion assembly in a temperature-sensitive mutant of Rous sarcoma virus.

A temperature-sensitive coordinate mutant tsLA83 of Prague (PR-B) strain of Rous sarcoma virus at the nonpermissive temperature (41 degrees) produces noninfectious virus particles (NI-LA83) which contained only 3% of the reverse-transcriptase activity present in infectious virions. Analyses of [35S]methionine-labeled NI-LA83 showed the presence of all of the viral proteins except reverse transcriptase. Pulse-chase analyses of the virus-specified proteins in cells infected with LA83 or PR-B showed that the gag and glycoprotein precursors, Pr76gag and gPr95env, respectively, were processed at both 35 and 41 degrees. The reverse-transcriptase precursor, Pr180gag-pol, however, was not processed in LA83-infected cells at 41 degrees. In contrast, cells infected with LA83 or PR-B at 35 degrees as well as with PR-B at 41 degrees showed normal cleavage of Pr180gag-pol. A shiftdown of LA83-infected cells at 41 degrees to the permissive temperature 35 degrees resulted in the normal processing of Pr180gag-pol and production of infectious virus containing reverse transcriptase. Electron microscopic analysis showed that at 41 degrees cells infected with LA83 showed a large number of budding structures but fewer released particles. A shiftdown from 41 to 35 degrees resulted in an increase of virus particles with a concomitant decrease in budding structures suggesting that the processing of reverse-transcriptase precursor is related to virion assembly.

Animals↗
PubMed1986

Polyproteins containing a domain encoded by the V-SKI oncogene are located in the nuclei of SKV-transformed cells.

SKV-transformed nonproducer clones were isolated from infected quail and chicken embryo cells. Analysis of intracellular viral RNAs by the Northern technique revealed that each clone contained a single SKV genome (either 5.7 or 8.9 kb) but no genome of the helper virus. Analysis of intracellular viral proteins containing gag determinants revealed that each clone contained a single species of either 55, 110, or 125 kDa. The intracellular location of these proteins was determined by indirect immunofluorescence employing either monoclonal antibodies (anti-p19gag) or conventional antiserum against gag proteins. All three of the SKV-specific proteins were localized to the nuclei of the transformed cells.

Animals↗
PubMed1986

Cytoskeletal association of an aphthovirus-induced polypeptide derived from the P3ABC region of the viral polyprotein.

Monolayers of BHK cells infected with aphthovirus (FMDV) were labeled for short times with [35S]methionine at 2 hr p.i. and fractionated by detergent treatment and low speed centrifugation. Polyacrylamide gel analysis showed an asymmetric distribution of the FMDV-induced polypeptides among the three different subcellular fractions obtained. Polypeptide P88-1, the viral capsid protein precursor, is mainly found in the soluble cytoplasmic extract while polypeptides P100-3, P52-2AC, P34-2C, and P14-2A are the major viral components of a detergent soluble extract of the crude nuclear pellet. Analysis of the detergent resistant fraction (DRF), which is mainly composed of cell nuclear chromatin and insoluble cytoskeletal elements, shows a clear enrichment in an incompletely characterized polypeptide which is tentatively designated P54. Variable amounts of polypeptides P100-3 and those of the P72 complex are also detected in this fraction. The preferential location of P54 in an equivalent subcellular fraction obtained by mild detergent treatment of infected monolayers in situ, and also in a high-salt resistant subfraction of the DRF, strongly suggests a close association of this polypeptide with vimentin-actin containing components of the cell. Polypeptide P54 is immunoprecipitated by viral specific antiserum from convalescent guinea pigs but not by serum against FMDV capsid proteins, indicating that it does not share common antigenic determinants with polypeptides processed from the viral capsid precursors. On the other hand, protease V8 mapping of polypeptides P100-3, P54, P88-1, and VP1-3 shows that P54 derived from the 3' end coding region of the viral genome. Further analysis by limited protease digestion also demonstrates that P54 has partial overlap with P72-3CD while it does not share any common peptide with P56a-3D, indicating that P54 contains the sequences coded in the 3ABC region of the FMDV RNA. This assumption is reinforced by the basic behavior shown by P54 in two-dimensional gels. The results support the hypothesis of a close intracellular interaction of a short-lived polypeptide, containing the viral protease and VPg sequences, with the host cytoskeleton, during infection of BHK cells with FMDV.

Animals↗
PubMed1988

Sindbis virus infection decreases intracellular pH: alkaline medium inhibits processing of Sindbis virus polyproteins.

Infection of baby hamster kidney cells by Sindbis virus, an alphavirus, resulted in a decrease in the intracellular pH of approximately 0.5 units within the first 1-2 hr after infection as measured either by equilibrium labeling with [14C]benzoic acid or by use of a pH-sensitive fluorescent probe, 2,7-bis-carboxyethyl-5,6-carboxyfluorescein-acetooxymethyl ester. In contrast, intralysosomal pH, as measured using an endocytized pH-sensitive probe, fluorescein isothiocyanate-labeled dextran, was not altered by Sindbis virus infection. Production of Sindbis virus was reduced by more than 90% and post-translational processing of Sindbis virus envelope precursors was inhibited in infected cells incubated in alkaline medium.

Animals↗
PubMed1988

Proteolytic processing of poliovirus polyprotein: elimination of 2Apro-mediated, alternative cleavage of polypeptide 3CD by in vitro mutagenesis.

The polypeptide 3CD of many poliovirus strains can be cleaved at two different amino acid pairs. The viral proteinase 3C and the viral polymerase 3D result from cleavage at a Gln-Gly pair by proteinase 3C, whereas cleavage at a Tyr-Gly pair by proteinase 2A yields the alternative products 3C' and 3D'. Specific mutations were introduced into the 3C'/3D' cleavage site in an infectious cDNA clone of poliovirus type 1 (Mahoney) by oligonucleotide-directed mutagenesis in order to investigate the role of 3C' and 3D' in viral proliferation and to obtain information about the cleavage specificity of 2Apro. Substitution of a threonine residue by an alanine residue at position -2 (P2) of this cleavage site abolished cleavage, whereas substitution of a tyrosine residue by a phenylalanine residue at amino acid position -1 (P1) of the cleavage site did not influence processing. Both mutated cDNA clones produced infectious viruses (T147A and Y148F) on transfection. The phenotypes of the mutant viruses were similar to that of the parental strain. We conclude that (i) 3C' and 3D' are not essential for virus replication, (ii) a Phe-Gly pair at the cleavage site can be cleaved by 2Apro, and (iii) a threonine residue in the P2 position of the cleavage site may be important in substrate recognition by 2Apro.

Amino Acid Sequence↗
PubMed1989

Specificity of enzyme-substrate interactions in foot-and-mouth disease virus polyprotein processing.

A series of transcripts derived from FMDV cDNA plasmids containing defined regions of the genome were translated in a rabbit reticulocyte lysate system. The products were analysed directly or following incubation with an FMDV-infected cell processing extract. Processing by the L proteinase at the L/1A cleavage site occurred when most of the P1-2A protein was absent. Substitution of sequences upstream of the 2C/3A cleavage site showed that the 3C proteinase was also able to cleave at an entirely novel cleavage site, apparently at K-I amino acid pairs. Cleavage at the 2A/2B site was not only independent of L and 3C proteinases, but was shown to occur when 2A and as few as four 2B N-terminal amino acids were present. Thus, the disparate proteolytic activities responsible for all three primary processing events that give rise to the products L, P1-2A, 2BC, and P3 were highly resistant either to major deletion or substitution of protein sequences adjacent to, or at, the site of cleavage. By contrast, secondary processing in trans was sensitive to changes at remote sites. For example, removal of the C-terminal regions of P1-2A and 2BC precursors impaired their ability to act as substrates for 3C proteinase activity. Processing of P1-2A, particularly of the 1D/2A cleavage site, was enhanced by inclusion of sequences from the 3D region of the genome.

Aphthovirus↗
PubMed1990

Cell-free translational analysis of the processing of infectious pancreatic necrosis virus polyprotein.

A cDNA clone of the large genomic segment of infectious pancreatic necrosis virus (IPNV) was inserted into transcription vectors and used for production of RNA transcripts of various lengths. These RNA transcripts were used to prime the synthesis of virus-specific polypeptides in a rabbit reticulocyte translation system. Full-length transcripts resulted in the synthesis of processed viral proteins pVP2, NS, and VP3. Transcripts which were deleted in the VP2 coding region did not affect the processing of NS or VP3 and similarly, deletions in VP3 did not affect the formation of NS or VP2. However, deletions which extended into the NS coding region resulted in a loss of protease activity and the production of truncated precursor polypeptides. The virus-specific proteolytic activity could not be inhibited by specific antisera and a trans activity for the viral proteases could not be demonstrated.

Animals↗
PubMed1992

Processing of VPg-containing polyproteins encoded by the B-RNA from cowpea mosaic virus.

To study the processing of putative VPg precursors the expression of specific mutant transcripts derived from a full-length cDNA clone of cowpea mosaic virus (CPMV) B-RNA was examined in a rabbit reticulocyte lysate system. This study revealed that the 170K protein produced by a B-RNA mutant that lacks the 32K coding region was efficiently processed by mainly intramolecular cleavages at three different sites into three sets of proteins of 60K + 110K, 84K + 87K, and 58K + 112K. Further cleavage of the 60K protein into 58K and VPg has not been observed in this in vitro system. The 84K protein can be further processed by an intramolecular cleavage reaction via two alternative pathways, either into 26K (VPg + 24K) and 58K proteins or into 24K and 60K proteins. VPg can be released from the 112K (VPg + 110K) precursor either directly or via the 26K intermediate. Immunoblot analysis showed that the 112K protein is present in CPMV-infected plant cells indicating that the in vitro observations may hold true in vivo.

Base Sequence↗
PubMed1992

Sequence upstream of the 24K protease enhances cleavage of the cowpea mosaic virus B RNA-encoded polyprotein at the junction between the 24K and 87K proteins.

To investigate cleavage at the junction between the cowpea mosaic virus (CPMV) 24K and 87K proteins, plasmids were constructed containing the sequence of bottom-component (B) RNA encoding the 110K protein plus a variable length of upstream coding sequence. Transcripts derived from these clones were translated in rabbit reticulocyte lysate and the appearance of the 87K protein was used to assess the efficiency of cleavage at the 24K-87K junction. The results show that the 110K protein, containing the contiguous sequence of the 24K and 87K proteins, is stable and that efficient cleavage at 24K-87K junction requires the presence of amino acids upstream of the 24K protease. These observations show that the 170K protein rather than the 110K protein is the precursor of the 87K protein and suggest a mechanism whereby both the B RNA-encoded 110K and 87K proteins can accumulate during infection.

Base Sequence↗
PubMed1993

The polyprotein allergens of nematodes.

Symptoms of infection with a number of parasitic nematodes may be associated with IgE antibody and associated immunopathologies. This has drawn attention to parasite allergens and provoked discussion on the wisdom of their inclusion in recombinant vaccines. Here, Larry McReynolds, Malcolm Kennedy and Murray Selkirk review work on a prominent set of allergens recently characterized in several nematode species.

Journal Article↗