A5074600548- Viral Infections and Immunology Research
- Animal Disease Management and Epidemiology
- RNA and protein synthesis mechanisms
- Viral Infectious Diseases and Gene Expression in Insects
- Virus-based gene therapy research
- Plant Virus Research Studies
- Viral gastroenteritis research and epidemiology
- Animal Virus Infections Studies
- CRISPR and Genetic Engineering
- Transgenic Plants and Applications
- Vector-Borne Animal Diseases
- RNA Interference and Gene Delivery
- Insect Resistance and Genetics
- Bacteriophages and microbial interactions
- Herpesvirus Infections and Treatments
- RNA modifications and cancer
- Peptidase Inhibition and Analysis
- Plant and Fungal Interactions Research
- Plant tissue culture and regeneration
- Chemical Synthesis and Analysis
- Monoclonal and Polyclonal Antibodies Research
- RNA regulation and disease
- Mosquito-borne diseases and control
- Insect-Plant Interactions and Control
- RNA Research and Splicing
University of St Andrews
2012-2024
Boston Children's Hospital
2013
The London College
2011
University of the West of England
2011
University of Oxford
1976-2011
University of Copenhagen
2011
University of Birmingham
2011
University of Alaska Anchorage
2011
University of Manchester
2011
Newcastle University
2011
The 2A region of the aphthovirus foot-and-mouth disease virus (FMDV) polyprotein is only 18 aa long. A ‘primary’ intramolecular processing event mediated by occurs at its own C terminus. FMDV activity was studied in artificial polyproteins which sequences encoding reporter proteins flanked sequence such that a single, long, open reading frame created. self-processing properties these were investigated and co-translational ‘cleavage’ products quantified. from our systems showed molar excess...
The 2A/2B cleavage of aphtho- and cardiovirus 2A polyproteins is mediated by their proteins 'cleaving' at own C termini. We have analysed this activity using artificial reporter polyprotein systems comprising green fluorescent protein (GFP) linked via foot-and-mouth disease virus (FMDV) to beta-glucuronidase (GUS) -- forming a single, long, open reading frame. Analysis the distribution radiolabel showed high proportion in vitro translation products (approximately 90%) were form 'cleavage'...
The 2A region of the foot-and-mouth disease virus (FMDV) polyprotein is only 16 amino acids in length. During synthesis FMDV a primary proteolytic processing event occurs between and 2B regions polyprotein. activity responsible for this cleavage not known but it thought that either an unidentified virus-encoded proteinase may be responsible, or acts as substrate host cell proteinase. A series recombinant polyproteins has been constructed which sequences to N- C-terminal side have deleted....
Mutating RNA virus genomes to alter codon pair (CP) frequencies and reduce translation efficiency has been advocated as a method generate safe, attenuated vaccines. However, selection for disfavoured CPs leads unintended increases in CpG UpA dinucleotide that also attenuate replication. We designed phenotypically characterised mutants of the picornavirus, echovirus 7, which these parameters were independently varied determine most influenced primarily replication ability while no fitness...
"2A" oligopeptides are autonomous elements containing a D(V/I)EXNPGP motif at the C terminus. Protein synthesis from an open reading frame internal 2A coding sequence yields two separate polypeptides, corresponding to sequences up and including those downstream. We show that reaction occurs in ribosomal peptidyltransferase center. Ribosomes pause end of sequence, over glycine proline codons, nascent chain this is released. Translation-terminating release factors eRF1 eRF3 play key roles...
Expression of viral proteins frequently includes non-canonical decoding events (‘recoding’) during translation. ‘2A’ oligopeptides drive one such event, termed ‘stop-carry on’ recoding. Nascent 2A peptides interact with the ribosomal exit tunnel to dictate an unusual stop codon-independent termination translation at final Pro codon 2A. Subsequently, ‘reinitiates’ on same codon, two individual being generated from open reading frame. Many have been identified, and they a conserved C-terminal...
Abstract The proliferation, differentiation, and survival of cells the mononuclear phagocyte system (MPS; progenitors, monocytes, macrophages, classical dendritic cells) are controlled by signals from M-CSF receptor (CSF1R). Cells MPS lineage have been identified using numerous surface markers transgenic reporters, but none is both universal restricted. In this article, we report development characterization a CSF1R reporter mouse. A FusionRed (FRed) cassette was inserted in-frame with C...
The primary 2A/2B polyprotein cleavage of aphtho-and cardioviruses is mediated by their 2A proteins cleaving C-terminally. Whilst the aphthovirus region only 16 aa (possibly 18 aa) long, cardiovirus protein some 150 aa. We have previously shown that foot-and-mouth disease virus (FMDV) able to mediate in an artificial (chloramphenicol acetyltransferase/FMDV 2A/beta-glucuronidase [CAT-2A-GUS]) system devoid any other FMDV sequences with high (approximately 85%), although not complete,...
During co-translational protein import into the endoplasmic reticulum ribosomes are docked onto translocon. This prevents inappropriate exposure of nascent chains to cytosol and, conversely, cytosolic factors from gaining access chain. We exploited this property translocation examine mechanism polypeptide cleavage by 2A peptide foot-and-mouth disease virus. find that scission reaction is unaffected placing a co-translationally targeted protein. Moreover, portion C-terminal site remains in...
2A is an oligopeptide sequence mediating a ribosome 'skipping' effect, producing apparent 'cleavage' of polyproteins. First identified and characterized in picornaviruses, '2A-like' sequences are found other mammalian viruses wide range insect viruses. Databases were analysed using motif conserved amongst 2A/2A-like sequences. The newly 2A-like (30 aa) inserted into reporter polyprotein to determine their cleavage activity. Our analyses showed that these fall two categories. majority...
ABSTRACT Infection of cells by picornaviruses leads to the generation intracellular membrane vesicles. The expression poliovirus (PV) 3A protein causes swelling endoplasmic reticulum (ER) and inhibition trafficking between ER Golgi apparatus. Here, we report that nonstructural proteins a second picornavirus, foot-and-mouth disease virus (FMDV), also perturb secretory pathway. FMDV 3A, 2B, 2C, 2BC expressed alone in were recovered from crude fractions, indicating association....
When a eukaryotic mRNA sequence specifying an amino acid motif known as 2A is directly followed by proline codon, two nonoverlapping proteins are synthesized. From earlier work, the second protein to start with this codon and not created proteolysis. Here we identify C-terminal of upstream 2A-encoded product from Perina nuda picorna-like virus that glycine specified last 2A-encoding sequence. This example recoding where promotes unconventional termination after decoding continued translation...
ABSTRACT Infection of cells with picornaviruses can lead to a block in protein secretion. For poliovirus this is achieved by the 3A protein, and consequent reduction secretion proinflammatory cytokines surface expression major histocompatibility complex class I proteins may inhibit host immune responses vivo. Foot-and-mouth disease virus (FMDV), another picornavirus, cause persistent infection ruminants, suggesting it too responses. Endoplasmic reticulum (ER)-to-Golgi apparatus transport...
The 18aa 2A self‐cleaving oligopeptide from foot‐and‐mouth disease virus can be used for co‐expression of multiple, discrete proteins a single ORF. mediates co‐translational cleavage at its own C‐terminus and is proposed to manipulate the ribosome into skipping synthesis specific peptide bond (producing discontinuity in backbone), rather than being involved proteolysis. To explore utility system target processing products, self‐processing polyproteins comprising fluorescent flanking were...
Where 2A oligopeptide sequences occur within ORFs, the formation of glycyl-prolyl peptide bond at C-terminus (each) does not occur. This property can be used to concatenate encoding several proteins into a single ORF: each component such an artificial polyprotein is generated as discrete translation product. and '2A-like' have become widely utilised in biotechnology biomedicine. Individual may also co- post-translationally targeted variety sub-cellular sites. In case polyproteins bearing...
Many biomedical applications absolutely require, or are substantially enhanced by, coexpression of multiple proteins from a single vector. Foot-and-mouth disease virus 2A (F2A) and “2A-like” sequences (e.g., Thosea asigna 2A; T2A) used widely for this purpose since can be coexpressed by linking open reading frames (ORFs) to form cistron. The activity F2A “cleavage” may, however, compromised both the use shorter versions (derived multiple-purpose cloning sites) link upstream protein. To...
Many biomedical applications require the expression or production of therapeutic hetero-multimeric proteins/protein complexes: in most cases only accomplished by co-ordinated co-expression within same cell. Foot-and-mouth disease virus 2A (F2A) and '2A-like' sequences are now widely used for this purpose. Since mediates a co-translational 'cleavage' at its own C-terminus, encoding multiple proteins (linked via 2As) can be concatenated into single ORF: transgene. It has been shown that some...
Foot-and-mouth disease virus (FMDV) is the type species of Aphthovirus genus Picornaviridae: Infection by picornaviruses results in a major rearrangement host cell membranes to create vesicular structures where genome replication takes place. In this report, using fluorescence and electron microscopy, membrane rearrangements cytoplasm FMDV-infected BHK-38 cells are documented. At 1.5-2.0 h post-infection, free ribosomes, fragmented rough endoplasmic reticulum, Golgi smooth membrane-bound...
Foot-and-mouth disease virus (FMDV) cDNA cassettes containing sequences encoding the capsid precursor P1-2A with and without those proteases L 3C were introduced into Autographa californica nuclear polyhedrosis (AcMNPV) expression vectors. Procapsid proteins 1AB, 1C 1D produced in cells infected recombinant baculoviruses, when present constructs, indicating that these FMDV active insect cells. Unlike P1 processing poliovirus, which has been shown to be catalysed mainly by 3CD gene product,...