Prions are transmissible agents caused by the self-propagating conformational change of proteins (32). Prions appear to be amyloid protein aggregates that propagate by capturing soluble proteins and converting them into an infectious aggregated form (33). According to the "protein only" hypothesis (32), the prion protein (PrP) is the sole agent responsible for causing numerous infectious diseases, including scrapie (sheep), bovine spongiform encephalopathy (cow), and chronic wasting disease (deer and elk) as well as kuru and Creutzfeld-Jacob disease (humans Het-s] in Podospora anserina, have also been characterized as non-Mendelian inheritable elements (7,37,43). Molecular and genetic studies of these fungal prions have greatly facilitated the elucidation of the molecular basis for prion conversion and propagation as well as the general criteria for prionogenicity in a protein's primary structure.[PSI ϩ ] is a prion form of Sup35, which is the eRF3 polypeptide release factor that is essential for terminating protein synthesis at stop codons (39, 45; for a review, see reference 17). When Sup35 is in the [PSI ϩ ] state, ribosomes often fail to release polypeptides at stop codons, causing a non-Mendelian trait to appear that is easily detected by nonsense suppression (23,29,30). To uncover host factors responsible for [PSI ϩ ] propagation, we have developed a genome-wide genetic selection method for [PSI ϩ ]-eliminating factors or mutants by use of the chromosomal ura3-197 mutant (21). Based on this selection system, we have selected host factors whose high-level expression on a multicopy plasmid leads to [PSI ϩ ] elimination. One clone yielded Rnq1⌬100, an N-terminal truncation of Rnq1, and is further examined in this study. Although there are some reports that the maintenance or de novo appearance of one prion is affected by several genetic manipulations such as overexpression of its own prion domain (15, 16), heterologous prion variants (5, 35), or nonprion protein mutants (1), the molecular basis of the action of one prion in inhibiting heterologous prions is not known.Rnq1 is a protein of unknown function and is one of several known yeast proteins containing a QN-rich prion domain, where the name derives from "rich in asparagine (N) and glutamine (Q)" (37 . According to the seeding model, a heterologous preexisting protein in the prion conformation is used as a template for the conversion of Sup35 into its prion form, which then proceeds to seed its own rapid and separate aggregation. Importantly, [PIN ϩ ] also facilitates the de novo appearance of the prion [URE3] and promotes polyglutamine (polyQ) aggregation and toxicity in general (5,25,27). Therefore, the seeding model predicts that [PIN ϩ ] aggregates provide a "friendly" nidus on which the first seeds of a heterologous prion or polyQ amyloid can form (11,41). The alternative titration model postulates that preexisting heterologous prions or prion-like aggregates capture and inactivate an inhibitor that prevents conversion of Sup35 into a
The Gpg1 protein is a G␥ subunit mimic implicated in the G-protein glucose-signaling pathway in Saccharomyces cerevisiae, and its function is largely unknown. Here we report that Gpg1 blocks the maintenance of [PSI ؉ ], an aggregated prion form of the translation termination factor Sup35. Although the GPG1 gene is normally not expressed, over-expression of GPG1 inhibits propagation of not only [PSI ؉ ] but also [PIN ؉ ], [URE3] prions, and the toxic polyglutamine aggregate in S. cerevisiae. Over-expression of Gpg1 does not affect expression and activity of Hsp104, a protein-remodeling factor required for prion propagation, showing that Gpg1 does not target Hsp104 directly. Nevertheless, prion elimination by Gpg1 is weakened by over-expression of Hsp104. Importantly, Gpg1 protein is prone to self-aggregate and transiently colocalized with Sup35NM-prion aggregates when expressed in [PSI ؉ ] cells. Genetic selection and characterization of loss-of-activity gpg1 mutations revealed that multiple mutations on the hydrophobic one-side surface of predicted ␣-helices of the Gpg1 protein hampered the activity. Prion elimination by Gpg1 is unaffected in the gpa2⌬ and gpb1⌬ strains lacking the supposed physiological G-protein partners of Gpg1. These findings suggest a general inhibitory interaction of the Gpg1 protein with other transmissible and nontransmissible amyloids, resulting in prion elimination. Assuming the ability of Gpg1 to form G-protein heterotrimeric complexes, Gpg1 is likely to play a versatile function of reversing the prion state and modulating the G-protein signaling pathway.
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