Identification of a Conserved Residue of Foamy Virus Gag Required for Intracellular Capsid Assembly

In contrast to all retroviruses but similar to the hepatitis B virus, foamy viruses (FV) require expression of the envelope protein for budding of intracellular capsids from the cell, suggesting a specific interaction between the Gag and Env proteins. Capsid assembly occurs in the cytoplasm of infected cells in a manner similar to that for the B- and D-type viruses; however, in contrast to these retroviruses, FV Gag lacks an N-terminal myristylation signal and capsids are not targeted to the plasma membrane (PM). We have found that mutation of an absolutely conserved arginine (Arg) residue at position 50 to alanine (R50A) of the simian foamy virus SFV cpz(hu) inhibits proper capsid assembly and abolishes viral budding even in the presence of the envelope (Env) glycoproteins. Particle assembly and extracellular release of virus can be restored to this mutant with the addition of an N-terminal Src myristylation signal (Myr-R50A), presumably by providing an alternate site for assembly to occur at the PM. In addition, the strict requirement of Env expression for capsid budding can be bypassed by addition of a PM-targeting signal to Gag. These results suggest that intracellular capsid assembly may be mediated by a signal akin to the cytoplasmic targeting and retention signal CTRS found in Mason-Pfizer monkey virus and that FV Gag has the inherent ability to assemble capsids at multiple sites like conventional retroviruses. The necessity of Env expression for particle egress is most probably due to the lack of a membrane-targeting signal within FV Gag to direct capsids to the PM for release and indicates that Gag-Env interactions are essential to drive particle budding.     

Resistance to a protein farnesyltransferase inhibitor in Plasmodium falciparum

The post-translational farnesylation of proteins serves to anchor a subset of intracellular proteins to membranes in eukaryotic organisms and also promotes protein-protein interactions. Inhibition of protein farnesyltransferase (PFT) is lethal to the pathogenic protozoa Plasmodium falciparum. Parasites were isolated that were resistant to BMS-388891, a tetrahydroquinoline (THQ) PFT inhibitor. Resistance was associated with a 12-fold decrease in drug susceptibility. Genotypic analysis revealed a single point mutation in the beta subunit in resistant parasites. The resultant tyrosine 837 to cysteine alteration in the beta subunit corresponded to the binding site for the THQ and peptide substrate. Biochemical analysis of Y837C-PFT demonstrated a 13-fold increase in BMS-388891 concentration necessary for inhibiting 50% of the enzyme activity. These data are consistent with PFT as the target of BMS-388891 in P. falciparum and suggest that PFT inhibitors should be combined with other antimalarial agents for effective therapy.     

Analysis of peptides synthesized in the presence of SAz-1 montmorillonite and Cu(2+) exchanged hectorite.

We have investigated the synthesis of oligopeptides containing glycine and tyrosine in the presence of the clay minerals montmorillonite (non-exchanged, SAz-1) and Cu(2+) exchanged hectorite. In both cases, homopolymers of the two amino acids are formed, as are mixed peptides. In the case of Cu(2+) hectorite, mixed oligopeptides up to trimers are detected in small amounts. For montmorillonite, heterogeneous oligopeptides up to hexamers are detected. Our experiments indicate montmorillonite is more effective in promoting oligopeptide formation than Cu(2+) hectorite. Analysis of the oligopeptide sequences formed on the montmorillonite surfaces indicates preferential synthesis of certain Gly-Tyr sequences over others.     

Biogeography and adaptation of Notothenioid fish: hemoglobin function and globin-gene evolution

The recognition of the important role of the polar habitats in global climate changes has awakened great interest in the evolutionary biology of polar organisms. They are exposed to strong environmental constraints, and it is important to understand how they have adapted to cope with these challenges and to what extent adaptations may be upset by current climate changes. We present an introductory overview of the evolution of the Antarctic fish fauna with emphasis on the dominant perciform sub-order Notothenioidei, as well as some specific comments on the biogeography of the three phyletically basal notothenioid families. The wealth of information on the ecology and biodiversity of the species inhabiting high-Antarctic and sub-Antarctic regions provides a necessary framework for better understanding the origin, evolution and adaptation of this unique group of fish. Notothenioidei offer opportunities for identification of the biochemical characters or the physiological traits responsible for thermal adaptation. The availability of phylogenetically related taxa in a wide range of latitudes has allowed to look into the molecular bases of environmentally driven phenotypic gain and loss of function. In the process of cold adaptation, the evolutionary trend of notothenioids has produced unique specialisations, including modification of hematological characteristics, e.g. decreased amounts and multiplicity of hemoglobins. The Antarctic family Channichthyidae (the notothenioid crown group) is devoid of hemoglobin. This loss is related to a single deletional event removing all globin genes with the exception of the inactive 3' end of adult alpha-globin. In reviewing hemoglobin structure, function and phylogeny, the evolution of the fish Root effect is analysed in detail. Adaptation of the oxygen-transport system in notothenioids seems to be based on evolutionary changes involving levels of biological organisation higher than the structure of hemoglobin.