A Cytoplasmic Complex Mediates Specific mRNA Recognition and Localization in Yeast

Müller, Marisa; Heym, Roland G.; Mayer, Andreas; Kramer, Katharina; Schmid, Maria; Cramer, Patrick; Urlaub, Henning; Jansen, Ralf‐Peter; Niessing, Dierk

doi:10.1371/journal.pbio.1000611

Cited by 74 publications

(165 citation statements)

References 52 publications

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“…It was shown recently that She3p also contains a zipcode-binding activity that was mapped to a C-terminal portion of She3p (10). Using in silico analyses, we detected another "homology box," only ∼20 residues upstream of the LPGV motif (Fig.…”

Section: Discussionmentioning

confidence: 87%

“…Because this upstream homology box contains conserved basic and aromatic residues, which are known to be preferred contact residues in RNA-protein interactions (25), this upstream homology box is a candidate site for zipcode binding. Moreover, previously reported cross-linking experiments identified residues 334-340 on the N-terminal side of the homology box (10). Because She2p's binding site for zipcode has been mapped to the "waist" of the She2p tetramer (dashed black box in Fig.…”

Section: Discussionmentioning

confidence: 99%

“…3). For quantitative pull-down assay (16) between MBP-She3pep and She2p, She2p (200 μL, 0.5 mg/mL in buffer D) was titrated with varying amount of MBP-She3pep-bound amylose beads (10,20,25,30,35,40,80,160, and 320 μL; 2 mg/mL in buffer D) in a final volume of 520 μL. MBPShe3pep-bound amylose beads were incubated with She2p for 30 min and then spinned down for 5 min at 15,000 × g. Supernatant was collected for each sample, and concentration was estimated by UV 280 .…”

Section: Methodsmentioning

confidence: 99%

“…Instead, cargo binds via an "adaptor" protein, swi5p-dependent HO expression 3 protein (She3p) (7). Notably, the 425-residue-long She3p contains at least four binding modules that appear to be sequentially arranged along its primary structure (8)(9)(10)(11). Our recent composite crystal structure of a region of She3p (comprising residues showed that two She3p molecules form a pseudocoiled coil (6).…”

mentioning

confidence: 99%

“…In addition to these two N-terminal binding sites, the C-terminal 92 residues of She3p are dedicated to interact with zipcode and She2p (Fig. 1A) (10). However, these sites have not yet been characterized at the atomic level, either in isolation or in complex with She2p and zipcode RNA.…”

mentioning

confidence: 99%

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Hooking She3p onto She2p for myosin-mediated cytoplasmic mRNA transport

Singh

Blobel

Shi

2014

Proc. Natl. Acad. Sci. U.S.A.

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The segregation of approximately two dozen distinct mRNAs from yeast mother to daughter cell cytoplasm is a classical paradigm for eukaryotic mRNA transport. The information for transport resides in an mRNA element 40–100 nt in length, known as “zipcode.” Targeted transport requires properly positioned actin filaments and cooperative loading of mRNA cargo to myosin. Cargo loading to myosin uses myosin 4 protein (Myo4p), swi5p-dependent HO expression 2 protein (She2p) and 3 protein (She3p), and zipcode. We previously determined a crystal structure of Myo4p and She3p, their 1:2 stoichiometry and interactome; we furthermore showed that the motor complex assembly requires two Myo4p⋅She3p heterotrimers, one She2p tetramer, and at least a single zipcode to yield a stable complex of [Myo4p⋅She3p⋅She2p⋅zipcode] in 2:4:4:1 stoichiometry in vitro. Here, we report a structure at 2.8-Å resolution of a cocrystal of a She2p tetramer bound to a segment of She3p. In this crystal structure, the She3p segment forms a striking hook that binds to a shallow hydrophobic pocket on the surface of each She2p subunit of the tetramer. Both She3p hook and cognate She2p binding pocket are composed of highly conserved residues. We also discovered a highly conserved region of She3p upstream of its hook region. Because this region consists of basic and aromatic residues, it likely represents part of She3p’s binding activity for zipcode. Because She2p also exhibits zipcode-binding activity, we suggest that “hooking” She3p onto She2p aligns each of their zipcode-binding activities into a high-affinity site, thereby linking motor assembly to zipcode.

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Section: Discussionmentioning

confidence: 87%

Section: Discussionmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

mentioning

confidence: 99%

mentioning

confidence: 99%

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Hooking She3p onto She2p for myosin-mediated cytoplasmic mRNA transport

Singh

Blobel

Shi

2014

Proc. Natl. Acad. Sci. U.S.A.

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Intracellular RNA Localization and Localized Translation

Besse

2013

Encyclopedia of Molecular Cell Biology and Molecular Medicine

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mTOR inhibition by rapamycin protects against deltamethrin‐induced apoptosis in PC12 Cells

Park

et al. 2015

Environmental Toxicology

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The autophagy pathway can be induced and upregulated in response to intracellular reactive oxygen species (ROS). In this study, we explored a novel pharmacotherapeutic approach involving the regulation of autophagy to prevent deltamethrin (DLM) neurotoxicity. We found that DLM-induced apoptosis in PC12 cells, as demonstrated by the activation of caspase-3 and -9 and by nuclear condensation. DLM treatment significantly decreased dopamine (DA) levels in PC12 cells. In addition, we observed that cells treated with DLM underwent autophagic cell death, by monitoring the expression of LC3-II, p62, and Beclin-1. Exposure of PC12 cells to DLM led to the production of ROS. Treatment with N-acetyl cysteine (NAC) effectively blocked both apoptosis and autophagy. In addition, mitogen-activated protein kinase (MAPK) inhibitors attenuated apoptosis as well as autophagic cell death. We also investigated the modulation of DLM-induced apoptosis in response to autophagy regulation. Pretreatment with the autophagy inducer, rapamycin, significantly enhanced the viability of DLM-exposed cells, and this enhancement of cell viability was partially due to alleviation of DLM-induced apoptosis via a decrease in levels of cleaved caspase-3. However, pretreatment of cells with the autophagy inhibitor, 3-methyladenine (3MA), significantly increased DLM toxicity in these cells. Our results suggest that DLM-induced cytotoxicity is modified by autophagy regulation and that rapamycin protects against DLM-induced apoptosis by enhancing autophagy. Pharmacologic induction of autophagy by rapamycin may be a useful treatment strategy in neurodegenerative disorders. © 2015 Wiley Periodicals, Inc. Environ Toxicol 32: 109-121, 2017.

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A Cytoplasmic Complex Mediates Specific mRNA Recognition and Localization in Yeast

Abstract: The localization of ash mRNA in yeast requires the binding of She2p and the myosin adaptor protein She3p to its localization element, which is highly specific and leads to the assembly of stable transport complexes.

Cited by 74 publications

References 52 publications

Hooking She3p onto She2p for myosin-mediated cytoplasmic mRNA transport

Hooking She3p onto She2p for myosin-mediated cytoplasmic mRNA transport

Intracellular RNA Localization and Localized Translation

mTOR inhibition by rapamycin protects against deltamethrin‐induced apoptosis in PC12 Cells

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