Identification of new genes that affect [PSI +] prion toxicity in Saccharomyces cerevisiae yeast

Matveenko, Andrew G.; Белоусов, М. В.; Bondarev, Stanislav A.; Moskalenko, Svetlana E.; Zhouravleva, Galina A.

doi:10.1134/s0026893316050113

Cited by 4 publications

(3 citation statements)

References 52 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…The 1-OT56 strain ( MAT a ade1–14 UGA his3 leu2 trp1–289 UAG ura3 [ psi − ][ PIN + ]) strain [41] of Saccharomyces cerevisiae was transformed with pRopA–YFP or pRopB–YFP plasmids for target proteins production or p315L–Cup1–YFP plasmid that was used as a negative control. Transformants were chosen on selective medium (SC) [42], and then single colonies were inoculated in liquid selective medium supplemented with 150 μM copper sulfate to induce the CUP1 promoter for overexpression of target proteins before growing for 6 h at 30 °C.…”

Section: Methodsmentioning

confidence: 99%

Two Novel Amyloid Proteins, RopA and RopB, from the Root Nodule Bacterium Rhizobium leguminosarum

et al. 2019

View full text Add to dashboard Cite

Amyloids represent protein fibrils with a highly ordered spatial structure, which not only cause dozens of incurable human and animal diseases but also play vital biological roles in Archaea, Bacteria, and Eukarya. Despite the fact that association of bacterial amyloids with microbial pathogenesis and infectious diseases is well known, there is a lack of information concerning the amyloids of symbiotic bacteria. In this study, using the previously developed proteomic method for screening and identification of amyloids (PSIA), we identified amyloidogenic proteins in the proteome of the root nodule bacterium Rhizobium leguminosarum. Among 54 proteins identified, we selected two proteins, RopA and RopB, which are predicted to have β-barrel structure and are likely to be involved in the control of plant-microbial symbiosis. We demonstrated that the full-length RopA and RopB form bona fide amyloid fibrils in vitro. In particular, these fibrils are β-sheet-rich, bind Thioflavin T (ThT), exhibit green birefringence upon staining with Congo Red (CR), and resist treatment with ionic detergents and proteases. The heterologously expressed RopA and RopB intracellularly aggregate in yeast and assemble into amyloid fibrils at the surface of Escherichia coli. The capsules of the R. leguminosarum cells bind CR, exhibit green birefringence, and contain fibrils of RopA and RopB in vivo.

show abstract

Section: Methodsmentioning

confidence: 99%

Two Novel Amyloid Proteins, RopA and RopB, from the Root Nodule Bacterium Rhizobium leguminosarum

et al. 2019

View full text Add to dashboard Cite

show abstract

“…Several attempts ta screening for such factors affecting the [PSI + ] prion or nonsense suppression have been made; however, these have yielded contradictory results. For example, SFP1 overexpression was found to enhance nonsense suppression on the [PSI + ] background, but not in other test systems [17,18,22,32]. Here, in order to uncover new mechanisms of translational accuracy, we concentrated on the effects of Sfp1 and Mcm1 on the transcription of the release factor genes.…”

Section: Discussionmentioning

confidence: 99%

“…We previously investigated the possible influence of two Q/N-rich transcription factors, Mcm1 and Sfp1, on [PSI + ] properties. Both were discovered by screening using a synthetic lethality test, which helped to identify factors that affect the lethality of [PSI + ] in the presence of sup45 mutation by influencing nonsense suppression [16][17][18]. Here, we attempted to find out whether the effects of Mcm1 and Sfp1 on [PSI + ] are due to their function as transcription regulators.…”

Section: Introductionmentioning

confidence: 99%

Transcription Factors Mcm1 and Sfp1 May Affect [PSI+] Prion Phenotype by Altering the Expression of the SUP35 Gene

Matveenko,

Mikhailichenko,

Drozdova

et al. 2024

Microbiology Research

View full text Add to dashboard Cite

Mcm1 is an essential Q/N-rich transcription factor. Q/N-rich proteins interact with each other, and many affect the [PSI+] prion formed by the translation termination factor Sup35 (eRF3). We found that transient MCM1 overexpression increased nonsense suppression in [PSI+] strains and SUP35 transcription. As we had discovered similar effects of another Q/N-rich transcription factor, Sfp1, here we focus on the roles of Mcm1 and Sfp1 in SUP35 expression, as well as on the effects of Sfp1 on the expression of the gene encoding another release factor, Sup45 (eRF1). Mutations in the SUP35 promoter showed that none of the potential Mcm1 binding sites affected the Sup35 protein level or nonsense suppression, even during MCM1 overexpression. Mcm1 itself neither formed aggregates in vivo nor affected Sup35 aggregation. In contrast, a mutation in the Sfp1-binding site decreased Sup35 production and [PSI+] toxicity of excess Sfp1. Mutation of the Sfp1 binding site in the SUP45 promoter lowered SUP45 expression and increased nonsense suppression even more drastically. Our data indicate that the mechanisms of Mcm1 and Sfp1 action differ. While Mcm1 seems unlikely to directly regulate SUP35 expression, Sfp1 appears to act through its binding sites and to directly activate SUP35 expression, which in turn may influence the [PSI+] prion phenotype and toxicity.

show abstract

The Gln3 Transcriptional Regulator of Saccharomyces cerevisiae Manifests Prion-Like Properties upon Overproduction

Antonets

Белоусов

Belousova

et al. 2019

Biochemistry Moscow

View full text Add to dashboard Cite

Identification of new genes that affect [PSI +] prion toxicity in Saccharomyces cerevisiae yeast

Cited by 4 publications

References 52 publications

Two Novel Amyloid Proteins, RopA and RopB, from the Root Nodule Bacterium Rhizobium leguminosarum

Two Novel Amyloid Proteins, RopA and RopB, from the Root Nodule Bacterium Rhizobium leguminosarum

Transcription Factors Mcm1 and Sfp1 May Affect [PSI+] Prion Phenotype by Altering the Expression of the SUP35 Gene

The Gln3 Transcriptional Regulator of Saccharomyces cerevisiae Manifests Prion-Like Properties upon Overproduction

Contact Info

Product

Resources

About