Hierarchical assembly of the Alu domain of the mammalian signal recognition particle

Weichenrieder, Oliver; Stehlin, Catherine; Kapp, U.; Birse, Darcy; Timmins, Peter; Strub, Katharina; Cusack, Stephen

doi:10.1017/s1355838201010160

Cited by 38 publications

(53 citation statements)

References 34 publications

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“…The likely monomeric physiological structure was inferred from the crystallographic model by proposing that the Alu RNA must fold back on to itself such that one SRP9/14 heterodimer interacts with a single RNA molecule, in a "closed" conformation ( Fig. 1D; Weichenrieder et al 2000Weichenrieder et al , 2001. This model was later confirmed by modeling into the low-resolution cryo-electron microscopy map of the entire mammalian SRP-ribosome complex (Halic et al 2004).…”

Section: Introductionmentioning

confidence: 85%

Section: Introductionmentioning

confidence: 99%

“…1A,B), but are otherwise mobile with respect to each other (Weichenrieder et al 2001). In previous work, we only succeeded in crystallizing this structure by restricting its flexibility, which was done by producing an artificial, circularly permuted Alu RNA with a rigid linkage (Weichenrieder et al 2000(Weichenrieder et al , 2001. The resulting Alu domain structure was a domain-swapped dimer with each RNA in the extended, "open" conformation and each SRP9/ 14 heterodimer bound to two sites, consistent with those mapped by hydroxyl radical cleavage experiments (Strub et al 1991), but on different adjacent RNA molecules ( Fig.…”

Section: Introductionmentioning

confidence: 99%

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Crystal structure of a signal recognition particle Alu domain in the elongation arrest conformation

Bousset¹,

Mary²,

Brooks³

et al. 2014

RNA

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The signal recognition particle (SRP) is a conserved ribonucleoprotein particle that targets membrane and secreted proteins to translocation channels in membranes. In eukaryotes, the Alu domain, which comprises the 5 ′ and 3 ′ extremities of the SRP RNA bound to the SRP9/14 heterodimer, is thought to interact with the ribosome to pause translation elongation during membrane docking. We present the 3.2 Å resolution crystal structure of a chimeric Alu domain, comprising Alu RNA from the archaeon Pyrococcus horikoshii bound to the human Alu binding proteins SRP9/14. The structure reveals how intricate tertiary interactions stabilize the RNA 5 ′ domain structure and how an extra, archaeal-specific, terminal stem helps constrain the Alu RNA into the active closed conformation. In this conformation, highly conserved noncanonical base pairs allow unusually tight side-by-side packing of 5 ′ and 3 ′ RNA stems within the SRP9/14 RNA binding surface. The biological relevance of this structure is confirmed by showing that a reconstituted full-length chimeric archaeal-human SRP is competent to elicit elongation arrest in vitro. The structure will be useful in refining our understanding of how the SRP Alu domain interacts with the ribosome.

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Section: Introductionmentioning

confidence: 85%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Crystal structure of a signal recognition particle Alu domain in the elongation arrest conformation

Bousset¹,

Mary²,

Brooks³

et al. 2014

RNA

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“…The SRP RNA might be involved, as it supports multiple conformations during the SRP cycle (35). The signal might act directly on the conformation of the Alu domain, which has been suggested to exist in two conformations (36). Alternatively, or in addition, flexible domains of the ribosome might be rearranged by long-range interactions within the ribosomal structure (37,38).…”

Section: Discussionmentioning

confidence: 99%

“…In addition, cross-linking experiments with SRP9/14 protein alone and with purified Alu RNPs gave apparently different cross-linked products than (42). The Alu domain is represented at scale by the SA50 Alu RNP structure (36). Its orientation was chosen randomly.…”

Section: Discussionmentioning

confidence: 99%

Signal Recognition Particle Alu Domain Occupies a Defined Site at the Ribosomal Subunit Interface upon Signal Sequence Recognition

et al. 2003

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The eukaryotic signal recognition particle (SRP) is essential for cotranslational targeting of proteins to the endoplasmic reticulum (ER). The SRP Alu domain is specifically required for delaying nascent chain elongation upon signal sequence recognition by SRP and was therefore proposed to interact directly with ribosomes. Using protein cross-linking, we provide experimental evidence that the Alu binding protein SRP14 is in close physical proximity of several ribosomal proteins in functional complexes. Crosslinking occurs even in the absence of a signal sequence in the nascent chain demonstrating that SRP can bind to all translating ribosomes and that close contacts between the Alu domain and the ribosome are independent of elongation arrest activity. Without a signal sequence, SRP14 cross-links predominantly to a protein of the large subunit. Upon signal sequence recognition, certain cross-linked products become detectable or more abundant revealing a change in the Alu domain-ribosome interface. At this stage, the Alu domain of SRP is located at the ribosomal subunit interface since SRP14 can be cross-linked to proteins from the large and small ribosomal subunits. Hence, these studies reveal differential modes of SRP-ribosome interactions mediated by the Alu domain.Ribosomes translating mRNAs coding for secretory and membrane proteins are specifically targeted to the endoplasmic reticulum (ER) membrane by a cytosolic ribonucleoprotein particle, the signal recognition particle (SRP) (for review, see ref 1). The specificity of this process is ensured by the presence of a signal sequence in the growing peptide chain, which is recognized by SRP when it emerges from the ribosome. Signal sequence recognition by SRP causes a slow down or an arrest in the elongation of the nascent chain (2, 3). The SRP-ribosome-nascent chain complex is then targeted to the ER via the interaction of SRP with its membrane receptor, the SRP receptor (SR), a heterodimeric membrane protein (4,5). SR coordinates the release of SRP from the ribosome with the insertion of the nascent chain into the Sec61 complex, the aqueous translocation pore in the ER membrane (for reviews, see refs 6 and 7). Free SRP can then engage in another targeting round, and membraneassociated ribosomes resume translation at their regular speed, leading to the cotranslational transfer of the nascent chain across or into the ER membrane. As emphasized by

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Entry into the Endoplasmic Reticulum: Protein Translocation, Folding and Quality Control

Fewell

Brodsky

2009

Trafficking Inside Cells

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Hierarchical assembly of the Alu domain of the mammalian signal recognition particle

Cited by 38 publications

References 34 publications

Crystal structure of a signal recognition particle Alu domain in the elongation arrest conformation

Crystal structure of a signal recognition particle Alu domain in the elongation arrest conformation

Signal Recognition Particle Alu Domain Occupies a Defined Site at the Ribosomal Subunit Interface upon Signal Sequence Recognition

Entry into the Endoplasmic Reticulum: Protein Translocation, Folding and Quality Control

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