Crystal structure of the human lysosomal mTORC1 scaffold complex and its impact on signaling

Araújo, Mariana E. G. de; Naschberger, Andreas; Fürnrohr, Barbara G.; Stasyk, Taras; Dunzendorfer-Matt, Theresia; Lechner, Stefan; Welti, Stefan; Kremser, Leopold; Shivalingaiah, Giridhar; Offterdinger, Martin; Lindner, Herbert; Huber, Lukas A.; Scheffzek, Klaus

doi:10.1126/science.aao1583

Cited by 118 publications

(106 citation statements)

References 59 publications

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“…The structure of Ragulator alone appears to be essentially the same as ours, reinforcing confidence in the accuracy of these structures. In the presence of the roadblock domains of RagA/C, Lamtor1 residues after 47 were found to be well ordered (de Araujo et al, 2017), consistent with our observation of HDX protection for Lamtor1 residues 61-70 in the presence of RagA/C. Interacting residues, such as Lamtor1 Val148, mutated in this study reduce function, consistent with expectations from our structure.…”

Section: Lamtor3supporting

confidence: 91%

“…In spite of the limited resolution of the EM structure, the orientation deduced for the RagA/C dimer relative to Ragulator is very similar. Similar conclusions were drawn based on the two different approaches applied, EM reconstruction with a full-length active RagA/C dimer in this case, and crystallization with the RagA/C roadblock fragments in the other study (de Araujo et al, 2017). The structural insights presented in these two studies will set the stage for a more detailed analysis of how the active Rag:Ragulator complex recruits and activates mTORC1, a central question at the heart of much current research into cellular metabolic regulation.…”

Section: Lamtor3supporting

confidence: 69%

“…While the initial version of this manuscript was under review, Scheffzek and colleagues reported the crystal structure of Ragulator alone and bound to the roadblock domains of RagA/C (de Araujo et al, 2017). The structure of Ragulator alone appears to be essentially the same as ours, reinforcing confidence in the accuracy of these structures.…”

Section: Lamtor3supporting

confidence: 66%

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Hybrid Structure of the RagA/C-Ragulator mTORC1 Activation Complex

Morris

Kim

et al. 2018

Biophysical Journal

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Section: Lamtor3supporting

confidence: 91%

Section: Lamtor3supporting

confidence: 69%

See 1 more Smart Citation

Hybrid Structure of the RagA/C-Ragulator mTORC1 Activation Complex

Morris

Kim

et al. 2018

Biophysical Journal

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“…At 16 Å, RagA and RagC appear essentially identical, and the assignment of RagA to one side and RagC to the other was ambiguous. On the basis of the report by de Araujo (2017), the assignment shown in Fig. 5C was selected.…”

Section: Resultsmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Hybrid Structure of the RagA/C-Ragulator mTORC1 Activation Complex

Morris

Kim

et al. 2017

Molecular Cell

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SUMMARY The lysosomal membrane is the locus for sensing cellular nutrient levels, which are transduced to mTORC1 via the Rag GTPases and the Ragulator complex. The crystal structure of the five-subunit human Ragulator at 1.4 Å resolution was determined. Lamtor1 wraps around the other four subunits to stabilize the assembly. The Lamtor2:Lamtor3 dimer stacks upon Lamtor4:Lamtor5 to create a platform for Rag binding. Hydrogen-deuterium exchange was used to map the Rag binding site to the outer face of the Lamtor2:Lamtor3 dimer and to the N-terminal intrinsically disordered region of Lamtor1. EM was used to reconstruct the assembly of the full-length RagAGTP:RagCGDP dimer bound to Ragulator at 16 Å resolution, revealing that the G-domains of the Rags project away from the Ragulator core. The combined structural model shows how Ragulator functions as a platform for the presentation of active Rags for mTORC1 recruitment, and might suggest an unconventional mechanism for Rag GEF activity.

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C7orf59/LAMTOR4 phosphorylation and structural flexibility modulate Ragulator assembly

et al. 2019

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Ragulator is a pentamer composed of p18, MP1, p14, C7orf59, and hepatitis B virus X‐interacting protein (HBXIP; LAMTOR 1—5) which acts as a lysosomal scaffold of the Rag GTPases in the amino acid sensitive branch of TORC1 signaling. Here, we present the crystal structure of human HBXIP‐C7orf59 dimer (LAMTOR 4/5) at 2.9 Å and identify a phosphorylation site on C7orf59 which modulates its interaction with p18. Additionally, we demonstrate the requirement of HBXIP‐C7orf59 to stabilize p18 and allow further binding of MP1‐p14. The structure of the dimer revealed an unfolded N terminus in C7orf59 (residues 1–15) which was shown to be essential for p18 binding. Full‐length p18 does not interact stably with MP1‐p14 in the absence of HBXIP‐C7orf59, but deletion of p18 residues 108–161 rescues MP1‐p14 binding. C7orf59 was phosphorylated by protein kinase A (PKA) in vitro and mutation of the conserved Ser67 residue to aspartate prevented phosphorylation and negatively affected the C7orf59 interaction with p18 both in cell culture and in vitro . C7orf59 Ser67 was phosphorylated in human embryonic kidney 293T cells. PKA activation with forskolin induced dissociation of p18 from C7orf59, which was prevented by the PKA inhibitor H‐89. Our results highlight the essential role of HBXIP‐C7orf59 dimer as a nucleator of pentameric Ragulator and support a sequential model of Ragulator assembly in which HBXIP‐C7orf59 binds and stabilizes p18 which allows subsequent binding of MP1‐p14.

show abstract

Crystal structure of the human lysosomal mTORC1 scaffold complex and its impact on signaling

Cited by 118 publications

References 59 publications

Hybrid Structure of the RagA/C-Ragulator mTORC1 Activation Complex

Hybrid Structure of the RagA/C-Ragulator mTORC1 Activation Complex

Hybrid Structure of the RagA/C-Ragulator mTORC1 Activation Complex

C7orf59/LAMTOR4 phosphorylation and structural flexibility modulate Ragulator assembly

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