Abstract The sobemoviruses have one of the smallest of all known RNA virus genomes. ORF1 encodes P1 which plays a role in suppression of silencing and virus movement, ORFs 2a and 2b encode the ...replicational polyproteins P2a and P2ab, and ORF3 encodes the coat protein. Translation of ORF2a from the genomic RNA is dependent on a leaky scanning mechanism. We report the presence of an additional ORF (ORFx), conserved in all sobemoviruses. ORFx overlaps the 5′ end of ORF2a in the +2 reading frame and also extends some distance upstream of ORF2a. ORFx lacks an AUG initiation codon and its expression is predicted to depend on low level initiation at near-cognate non-AUG codons, such as CUG, by a proportion of the ribosomes that are scanning the region between the ORF1 and ORF2a initiation codons. Mutations that disrupt translation of ORFx in turnip rosette virus prevent the establishment of infection.
Theiler's murine encephalomyelitis virus (TMEV) is a member of the genus Cardiovirus in the Picornaviridae, a family of positive-sense single-stranded RNA viruses. Previously, we demonstrated that in ...the related cardiovirus, Encephalomyocarditis virus, a programmed-1 ribosomal frameshift (1 PRF) occurs at a conserved G_GUU_UUU sequence within the 2B-encoding region of the polyprotein open reading frame (ORF). Here we show that-1 PRF occurs at a similar site during translation of the TMEV genome. In addition, we demonstrate that a predicted 3= RNA stem-loop structure at a noncanonical spacing downstream of the shift site is required for efficient frameshifting in TMEV and that frameshifting also requires virus infection. Mutating the G_GUU_UUU shift site to inhibit frameshifting results in an attenuated virus with reduced growth kinetics and a small-plaque phenotype. Frameshifting in the virus context was found to be extremely efficient at 74 to 82%, which, to our knowledge, is the highest frameshifting efficiency recorded to date for any virus. We propose that highly efficient-1 PRF in TMEV provides a mechanism to escape the confines of equimolar expression normally inherent in the single-polyprotein expression strategy of picornaviruses.
Atmospheric and oceanic partial pressures of CO2 (pCO2) have been recorded automatically along two Atlantic meridional transects in 1995. The tropical Atlantic ocean (20°S–20°N) is generally a source ...of CO2 for the atmosphere, but in the region of the North Equatorial Countercurrent an undersaturation of CO2 has been observed. Undersaturations previously reported in the literature are explained by the decrease of salinity due to the high precipitations associated with the Intertropical Convergence Zone. In June 1995, strong CO2 undersaturations (ΔpCO2 = −70μatm) were observed near 8°N, which suggests, in addition of the salinity effect, an uptake of CO2 due to biological activity. This undersaturation, although weaker than in spring, also appeared at other periods of the year 1995.
Highlights • Definition of Clostridium difficile toxin-derived antigens for soluble expression in E. coli. • Demonstration of their potent neutralising immune response against key epidemic strain ...toxins. • TcdA and TcdB were different with respect to the domains that evoke a neutralising immune response. • TcdB central domains dominate the generation of a toxin-neutralising response. • Generated antibodies prevent C. difficile infection in passive immunisation studies.
We describe mutations in the hepatitis B virus (HBV) polymerase gene in viruses which reactivated in two patients during therapy with -2'-deoxy- 3'-thiacytidine, or lamivudine (3TC), and following ...orthotopic liver transplantation for chronic hepatitis B. Virus resistance to 3TC is associated with mutations which lead to amino acid substitutions in the highly conserved tyr-met-asp-asp (YMDD) motif, part of the active site of the polymerase, and which parallel those seen in resistant human immunodeficiency virus (HIV). Substitutions of valine and isoleucine for methionine were found in the two cases. The significance of single secondary mutations, which differ between viruses from the two patients, remains to be determined. Thus, viral resistance to lamivudine of hepatitis B virus mimics that of HIV and can occur in the setting of immunosuppression after liver transplantations. (Hepatology 1996 Sep;24(3):711-3)