Analysis of stress granule assembly in <i>Schizosaccharomyces pombe</i>

Wang, Chunyu; Wen, Wei-Ling; Nilsson, Daniel; Sunnerhagen, Per; Wang, Shao-Win

doi:10.1261/rna.030270.111

Cited by 31 publications

(41 citation statements)

References 34 publications

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“…For example, we found that Ubp3 colocalizes with the stress granule marker Pbp1 in heatstressed cells. A similar association has been observed in S. pombe cells exposed to a heat stress (50). Furthermore, USP10 was observed to localize to stress granules in mammalian cells exposed to heat or arsenite stress (28).…”

Section: Discussionsupporting

confidence: 80%

The Catalytic Activity of the Ubp3 Deubiquitinating Protease Is Required for Efficient Stress Granule Assembly in Saccharomyces cerevisiae

Nostramo

Varia

Zhang

et al. 2016

Molecular and Cellular Biology

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The interior of the eukaryotic cell is a highly compartmentalized space containing both membrane-bound organelles and the recently identified nonmembranous ribonucleoprotein (RNP) granules. This study examines in Saccharomyces cerevisiae the assembly of one conserved type of the latter compartment, known as the stress granule. Stress granules form in response to particular environmental cues and have been linked to a variety of human diseases, including amyotrophic lateral sclerosis. To further our understanding of these structures, a candidate genetic screen was employed to identify regulators of stress granule assembly in quiescent cells. These studies identified a ubiquitin-specific protease, Ubp3, as having an essential role in the assembly of these RNP granules. This function was not shared by other members of the Ubp protease family and required Ubp3 catalytic activity as well as its interaction with the cofactor Bre5. Interestingly, the loss of stress granules was correlated with a decrease in the long-term survival of stationary-phase cells. This phenotype is similar to that observed in mutants defective for the formation of a related RNP complex, the Processing body. Altogether, these observations raise the interesting possibility of a general role for these types of cytoplasmic RNP granules in the survival of G 0 -like resting cells.T he interior of the eukaryotic cell is a highly compartmentalized space. Organelles harbor distinct sets of proteins that often possess related functions, serving to organize the cellular milieu and increase reaction efficiencies. The recent identification of a family of ribonucleoprotein (RNP) granules suggests that this complexity may be greater than previously recognized (1, 2). These RNP granules differ from traditional organelles in that they lack a limiting membrane and form in response to particular cues, including cellular stress and developmental signals (3). Interest in these structures has increased sharply as they have been recently linked to a number of human diseases, including neurodegenerative disorders like amyotrophic lateral sclerosis and spinocerebellar ataxia type II (4,5). Although the function of many of these granules is unknown, their conservation throughout evolution suggests that they serve an important biological role.Stress granules are one of the better-characterized instances of these cytoplasmic RNP complexes. They form in response to various types of cellular stress and in quiescent or G 0 cells (3, 6). They are highly dynamic structures, continuously shuttling components between the cytosol and the granule during the stress and disassembling rapidly following its cessation (7). In mammalian cells, stress granule formation is initiated by the translation arrest that occurs upon the stress-triggered phosphorylation of translation initiation factor 2␣ (eIF2␣) (8). Polysomes disassemble, and 48S preinitiation complexes containing naked mRNA accumulate and are bound by mRNA-binding proteins that contain low-complexity domains (8). These domains...

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Section: Discussionsupporting

confidence: 80%

The Catalytic Activity of the Ubp3 Deubiquitinating Protease Is Required for Efficient Stress Granule Assembly in Saccharomyces cerevisiae

Nostramo

Varia

Zhang

et al. 2016

Molecular and Cellular Biology

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“…Images were acquired using a Leica DM RA2 microscope equipped with a Leica DC 350F camera and were assembled using Adobe PhotoShop. Image quantitation was done as described previously (11).…”

Section: Methodsmentioning

confidence: 99%

“…The two-step immunoglobulin G (IgG)-Sepharose and calmodulin resin affinity purification was carried out as described previously (11). The purified proteins were eluted with 20 mM EGTA to be precipitated with trichloroacetic acid (TCA) at a final concentration of 20%, separated on SDS-PAGE gels, and visualized by Sypro Ruby.…”

Section: Methodsmentioning

confidence: 99%

Pdc1 Functions in the Assembly of P Bodies in Schizosaccharomyces pombe

Wang

Chen

Wang

2013

Molecular and Cellular Biology

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P bodies are cytoplasmic RNA granules containing the Dcp1-Dcp2 decapping enzymes where mRNA decay can occur. Here, we describe the characterization of P bodies in the fission yeast Schizosaccharomyces pombe. Most information on the property and function of P bodies stems from studies in the distantly related budding yeast Saccharomyces cerevisiae, and Edc3 was identified as a scaffold protein required for P-body assembly. However, we found that, unlike in S. cerevisiae, fission yeast Edc3 was dispensable for P-body formation. Pdc1, a novel partner of the fission yeast decapping enzyme, with a limited similarity to plant Edc4/Varicose that is required for the assembly of P bodies, was identified (tandem affinity purification-matrix-assisted laser desorption ionization tandem mass spectrometry [TAP-MALDI MS/MS]). Pdc1 interacts with Dcp2 through its C terminus and contains a coiled-coil region for self-interaction to mediate P-body formation. In line with the model that Pdc1 cross-bridges different proteins, additional interactions can be demonstrated with components such as Edc3 and Ste13. Although Pdc1 is not required for the interaction between Dcp1 and Dcp2, our data suggest that Pdc1 acts as a functional homologue of Edc4, a third component of the decapping enzymes that is thought to be absent from fungi. Together, these results highlight the diverse Pbody protein compositions between different species and might help to provide insight into their evolutionary paths. C ytoplasmic processing bodies (P bodies) are dynamic RNA protein aggregates that play critical roles in mRNA degradation, nonsense-mediated mRNA decay, translational repression, and RNA-mediated gene silencing (reviewed in reference 1). Because several components of P bodies, such as the decapping enzyme and its coactivators, are found in both yeast and mammalian cells, it is believed that P bodies are evolutionarily conserved among eukaryotes. However, a detailed study of P bodies in the fission yeast has not yet been described.The decapping of mRNAs, which occurs inside P bodies, represents a critical step in mRNA turnover and is therefore tightly regulated. The decapping enzyme Dcp2 requires additional proteins for full activity and/or stability. These proteins are generally termed decapping coactivators (or enhancers of decapping [EDCs]) although they may activate decapping by different mechanisms (2). In Saccharomyces cerevisiae, proteins that activate decapping include Dcp1, enhancer of decapping 1 to 3 (Edc1-3), the heptameric Lsm1-7 complex, DEXH/D-box RNA helicase 1 (Dhh1; RCK/p54 in mammals), and Pat1 (3). Dcp1 interacts directly with Dcp2 and is required for decapping in vivo (4). All of these proteins colocalize to P bodies.In human cells, the interaction between Dcp1 and Dcp2 appears to be mediated by the decapping activator Edc4 (also known as Ge-1 or Hedls, for human enhancer of decapping large subunit) (5, 6). Edc4, Dcp1, and Dcp2 are part of a multimeric protein complex, which also includes Edc3 and RCK/p54 (5). Similarly, Arabid...

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“…PABP is a marker of stress granules, RNA-enriched granules that are formed in response to certain stress conditions (38), whereas Hsp104 is a large molecular chaperone involved in protein quality control and disaggregation of various cytosolic protein aggregates (39). To further examine the nature of the observed Sdj1-L169P aggregates, the localization of Sdj1-L169P was analyzed in strains expressing mCherry-labeled PABP (Pab1) or Hsp104.…”

Section: Volume 290 • Number 34 • August 21 2015mentioning

confidence: 99%

A Two-step Protein Quality Control Pathway for a Misfolded DJ-1 Variant in Fission Yeast

Mathiassen

Larsen

Poulsen

et al. 2015

Journal of Biological Chemistry

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Background: A mutation, L166P, in DJ-1, is linked to Parkinson disease. Results: The Sdj1-L169P fission yeast orthologue of DJ1-L166P is misfolded, associated with chaperones, and degraded via two ubiquitin-proteasome dependent pathways. Conclusion: Sdj1-L169P is subject to a two-step degradation pathway. Significance: Mapping the degradation pathways for misfolded proteins is important for our basic understanding of protein quality control in health and disease.

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Analysis of stress granule assembly in Schizosaccharomyces pombe

Cited by 31 publications

References 34 publications

The Catalytic Activity of the Ubp3 Deubiquitinating Protease Is Required for Efficient Stress Granule Assembly in Saccharomyces cerevisiae

The Catalytic Activity of the Ubp3 Deubiquitinating Protease Is Required for Efficient Stress Granule Assembly in Saccharomyces cerevisiae

Pdc1 Functions in the Assembly of P Bodies in Schizosaccharomyces pombe

A Two-step Protein Quality Control Pathway for a Misfolded DJ-1 Variant in Fission Yeast

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