CONFLIT GÉNÉTIQUE (French Edition)
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Here we briefly describe how we developed a yeast model to identify drugs and host cell factors that interfere with the capacity of the Epstein-Barr virus EBV to evade the host immune system. EBV evades the host immune system but, fortunately, has an Achilles heel: its genome maintenance protein EBNA1 which is, on the one hand, essential for EBV genome replication and maintenance and as such expressed in all dividing EBV-infected cells. Hence, in order to avoid the immune system to detect and kill EBV-infected cells, the virus has evolved a mechanism to limit EBNA1 production to the minimal level to sustain viral genome replication and maintenance and, at the same time, to minimize the production of EBNA1-derived antigenic peptides.
This inhibitory effect of GAr is length-dependent as a longer domain displays a stronger inhibitory effect on both mRNA translation and antigen presentation. Of note, a polymorphism in the length of GAr exists, which may reflect a way for EBV to adapt to its host. GAr-based EBNA1 immune evasion has been considered a relevant therapeutic target to treat EBV-related cancers, hence there is a need to develop cell-based assays that allow high throughput screenings. As the mechanism of GAr-mediated mRNA translation suppression in cis was not known, nor the cellular factors involved, a yeast Saccharomyces cerevisiae assay that recapitulates all the aspects of the GAr-based inhibition of translation, including the GAr-length dependency, has been developed.
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Briefly, this assay is based on the construction of yeast strains that express individually fusions between GAr of various lengths and the Ade2p reporter protein as sole source of Ade2p enzyme. Yeast cells which express Ade2p at a functional level form white colonies, whereas cells that do not express Ade2p readily form red colonies and cells that express any intermediate level of Ade2p form pink colonies whose coloration intensity is inversely proportional to the level of Ade2p expressed.
Because GAr is also able to self-limit in a length-dependent manner the translation of its own mRNA in yeast, this leads to a reduction in Ade2p level which is proportional to GAr size and can be easily monitored by looking at the color of the yeast colonies. A yeast strain that endogenously expressed a fusion between a 43 amino acids GAr and Ade2p was chosen because it leads to a partial inhibition of Ade2p expression, thus leading to pink colonies expressing an intermediate level of Ade2p.
Hence, this 43GAr-ADE2 strain allows to look for modifiers drugs or cellular factors that either suppress or exacerbate the inhibitory effect of GAr on translation Fig.
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This 43GAr-ADE2 strain was successfully used to identify small molecular weight compounds that can stimulate 43GAr-Ade2p expression in yeast, hence leading to whiter colonies that express higher level of 43GAr-Ade2 Fig. This yeast model for EBNA1 stealthiness was validated since these drugs, which include doxorubicin, were later shown to also stimulate EBNA1 expression in EBV-infected human cells and to relieve GAr-based limitation of antigen presentation.
These results also suggested that potential host cell genes involved in EBV immune evasion were conserved in yeast.
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B Rationale of the yeast model for GAr-dependent inhibition of translation. As GAr is also operant in yeast, the 43GAr-Ade2p fusion protein is only weakly expressed in a strain where it is the only source of Ade2p, thereby leading to the formation of pink colonies that express a limited amount of the protein as compared to a control strain expressing a higher Ade2p level, thus forming white colonies. Hence, based on changes in the intensity of its pink color, this strain can be used to look for modifiers that either suppress or exacerbate the GAr inhibitory effect on translation.
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C This 43GAr strain was used for both drug and genetic screening, leading to the isolation of doxorubicin which leads to a GAr-dependent increase in translation and to nucleolin which is a host factor involved in the GAr inhibitory effect on translation. D The effect of both doxorubicin and nucleolin are GAr-dependent since they have no effect on the control strain expressing Ade2p.
Hence, Lista et al.
Indeed, when overexpressed in the pink 43GAr-ADE2 strain, Nsr1p led to the formation of redder yeast colonies that express even less 43GAr-Ade2p, whereas the knockout of the NSR1 gene which is viable in yeast in the same strain led to white colonies that express high level of 43GAr-Ade2p, showing that Nsr1p is involved in GAr-based limitation of protein expression Fig. This book describes a range of different mycoviruses and their geographical distribution, transmission and evolution, together with their effects on the fungal hosts and how these are brought about.
Keywords RNA silencing gemycircularvirus mycovirus antiviral dicer dsRNA mycoviruses multiplex PCR Aspergillus fumigatus chrysovirus Aspergillus fumigatus partitivirus-1 Aspergillus fumigatus tetramycovirus Satellite altimetry has provided extremely valuable information on ocean science e. With more than 25 years of observations, it is also becoming vital to climate research, providing accurate measurements of sea level variations from regional to global scales.
Altimetry has also demonstrated a strong potential for geophysical, cryospheric, and hydrological research and is now commonly used for the monitoring of Arctic and Antarctic ice sheet topography and of terrestrial surface water levels. This book aims to present reviews and recent advances of general interest in the use of radar altimetry in Earth sciences. Manuscripts are related to any aspect of radar altimetry technique or geophysical applications.
We also encourage manuscripts resulting from the application of new altimetric technology SAR, SARin, and Ka band and improvements expected from missions to be launched in the near future i. OpenEdition Licence for Books 5. CC by-nc-nd 2. OpenEdition licence for Books 2.
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