Cryo-EM structure of C9ORF72–SMCR8–WDR41 reveals the role as a GAP for Rab8a and Rab11a

Tang, Dan; Sheng, J.; Xu, Lianjun; Zhan, Xiechao; Liu, Jiaming; Jiang, Hui; Shu, Xi; Liu, Xiaoyu; Zhang, Tizhong; Jiang, Lan; Zhou, Caijin; Li, Wenqi; Cheng, Wei; Li, Zhonghan; Wang, Kunjie; Lu, Kefeng; Yan, Chuangye; Qi, Shiqian

doi:10.1073/pnas.2002110117

Cited by 63 publications

(79 citation statements)

References 50 publications

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“…The secondary structure predicted by WDSP, a database for WD40 proteins, also predicted a similar large loop extending from the WDR41 β-propeller (Ma et al, 2019; Y. Wang et al, 2015). Although not available at the start of our study, two recently reported cryo-EM structures for the WDR41-C9orf72-SMCR8 complex validate this prediction for the overall structural organization of WDR41 and showed that the interaction with SMCR8-C9orf72 is via the WDR41 β-propeller (23,24).…”

Section: Prediction Of a Novel Mechanism For The Wdr41-pqlc2 Interactionsupporting

confidence: 75%

Receptor-like role for PQLC2 amino acid transporter in the lysosomal sensing of cationic amino acids

Talaia

Amick

Ferguson

2020

Preprint

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PQLC2, a lysosomal cationic amino acid transporter, also serves as a sensor that responds to scarcity of its substrates by recruiting a protein complex comprised of C9orf72, SMCR8 and WDR41 to the surface of lysosomes. This protein complex controls multiple aspects of lysosome function. Although it is known that this response to changes in cationic amino acid availability depends on an interaction between PQLC2 and WDR41, the underlying mechanism for the regulated interaction is not known. In this study, we establish that the WDR41-PQLC2 interaction is mediated by a short peptide motif in a flexible loop that extends from the WDR41 β-propeller and inserts into a cavity presented by the inward-facing conformation of PQLC2. This data supports a transceptor model wherein conformational changes in PQLC2 related to substrate transport regulate the availability of the WDR41 binding site on PQLC2 and mediate recruitment of the WDR41-SMCR8-C9orf72 complex to the surface of lysosomes.

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Section: Prediction Of a Novel Mechanism For The Wdr41-pqlc2 Interactionsupporting

confidence: 75%

Receptor-like role for PQLC2 amino acid transporter in the lysosomal sensing of cationic amino acids

Talaia

Amick

Ferguson

2020

Preprint

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“…Farg et al [ 43 ] first reported C9orf72 to interact with RAB1, RAB5, RAB7 and RAB11. Webster et al [ 22 ] confirmed that C9orf72 associates with GTP-bound RAB1A and the ULK1 complex, and it has been demonstrated that C9orf72 in complex with SMCR8 and WDR41 is a GEF for RAB8A, RAB11A, and RAB39B, and that its loss perturbs autophagy in neurons [ 27 , 29 , 31 , 89 ]. We detected only RAB1B in our SMCR8 and C9orf72 interactomes (Tables S1 , S2 ), but failed to confirm binding of V5-tagged RAB1A, a paralog highly similar in sequence to RAB1A, with SMCR8 in direct co-IP experiments.…”

Section: Resultsmentioning

confidence: 99%

C9orf72-associated SMCR8 protein binds in the ubiquitin pathway and with proteins linked with neurological disease

Goodier

Soares

Pereira

et al. 2020

acta neuropathol commun

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A pathogenic GGGCCC hexanucleotide expansion in the first intron/promoter region of the C9orf72 gene is the most common mutation associated with amyotrophic lateral sclerosis (ALS). The C9orf72 gene product forms a complex with SMCR8 (Smith-Magenis Syndrome Chromosome Region, Candidate 8) and WDR41 (WD Repeat domain 41) proteins. Recent studies have indicated roles for the complex in autophagy regulation, vesicle trafficking, and immune response in transgenic mice, however a direct connection with ALS etiology remains unclear. With the aim of increasing understanding of the multi-functional C9orf72-SMCR8-WDR41 complex, we determined by mass spectrometry analysis the proteins that directly associate with SMCR8. SMCR8 protein binds many components of the ubiquitin-proteasome system, and we demonstrate its poly-ubiquitination without obvious degradation. Evidence is also presented for localization of endogenous SMCR8 protein to cytoplasmic stress granules. However, in several cell lines we failed to reproduce previous observations that C9orf72 protein enters these granules. SMCR8 protein associates with many products of genes associated with various Mendelian neurological disorders in addition to ALS, implicating SMCR8-containing complexes in a range of neuropathologies. We reinforce previous observations that SMCR8 and C9orf72 protein levels are positively linked, and now show in vivo that SMCR8 protein levels are greatly reduced in brain tissues of C9orf72 gene expansion carrier individuals. While further study is required, these data suggest that SMCR8 protein level might prove a useful biomarker for the C9orf72 expansion in ALS.

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“…mTORC1 negatively regulates autophagy, so the ARF1-mTORC1 connection could explain how haploinsufficient C9orf72 leads to a decrease in autophagy, which has in turn been linked to multiple neurodegenerative diseases 30 . While our paper was under review, a cryo-EM structure of a dimeric form of this complex was reported and proposed to serve as a GAP for RAB8A and RAB11A 31 . The relative roles of GAP activity with respect to different small GTPases in normal function and disease remain to be determined.…”

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confidence: 99%

Structure of the C9orf72 ARF GAP complex that is haploinsufficient in ALS and FTD

Fromm

Zoncu

et al. 2020

Nature

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Mutation of C9ORF72 is the most prevalent defect in amyotrophic lateral sclerosis (ALS) and frontal temporal degeneration (FTD). Together with hexanucleotide repeat expansion, haploinsufficiency of C9ORF72 contributes to neuronal dysfunction. We determined the structure of the SMCR8-C9orf72-WDR41 complex by cryo-EM. C9orf72 and SMCR8 are both longin-DENN domain proteins, while WDR41 is a beta-propeller protein that binds to SMCR8 such that the whole structure resembles an eye slip hook. Contacts between WDR41 and SMCR8 DENN drive lysosomal localization in amino acid starvation. The structure suggested that SMCR8-C9orf72 was a small GTPase activating protein (GAP). We found that SMCR8-C9orf72-WDR41 is a GAP for Arf family small GTPases, and refer to it as the Lysosomal SMCR8-C9orf72 Arf GAP ("L-SCARF") complex. These data rationalize the function of C9orf72 both in normal physiology and in ALS/FTD.

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Cryo-EM structure of C9ORF72–SMCR8–WDR41 reveals the role as a GAP for Rab8a and Rab11a

Cited by 63 publications

References 50 publications

Receptor-like role for PQLC2 amino acid transporter in the lysosomal sensing of cationic amino acids

Receptor-like role for PQLC2 amino acid transporter in the lysosomal sensing of cationic amino acids

C9orf72-associated SMCR8 protein binds in the ubiquitin pathway and with proteins linked with neurological disease

Structure of the C9orf72 ARF GAP complex that is haploinsufficient in ALS and FTD

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