Cryo-EM structure of the human ELMO1-DOCK5-Rac1 complex

Kukimoto-Niino, Mutsuko; Katsura, Kazushige; Kaushik, Rahul; Ehara, Haruhiko; Yokoyama, Takeshi; Uchikubo‐Kamo, Tomomi; Nakagawa, Reiko; Mishima-Tsumagari, Chiemi; Yonemochi, Mayumi; Ikeda, Mariko; Hanada, Kazuharu; Zhang, Kam Y. J.; Shirouzu, Mikako

doi:10.1126/sciadv.abg3147

Cited by 19 publications

(30 citation statements)

References 64 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…In this regard, it is noteworthy that activated Rac3 was previously identified as a NRBP1 binding partner that co-localized with NRBP1 at endomembranes and in lamellopodia [10]. Such a scaffolding function for NRBP1 would be similar to that characterized for ELMO1, which associates with both Rac1 and the Rac GEF DOCK5, and enhances the GEF activity of DOCK5 [34].…”

Section: Discussionmentioning

confidence: 81%

The pseudokinase NRBP1 activates Rac1/Cdc42 via P-Rex1 to drive oncogenic signalling in triple-negative breast cancer

et al. 2023

View full text Add to dashboard Cite

We have determined that expression of the pseudokinase NRBP1 positively associates with poor prognosis in triple negative breast cancer (TNBC) and is required for efficient migration, invasion and proliferation of TNBC cells in culture as well as growth of TNBC orthotopic xenografts and experimental metastasis. Application of BioID/MS profiling identified P-Rex1, a known guanine nucleotide exchange factor for Rac1, as a NRBP1 binding partner. Importantly, NRBP1 overexpression enhanced levels of GTP-bound Rac1 and Cdc42 in a P-Rex1-dependent manner, while NRBP1 knockdown reduced their activation. In addition, NRBP1 associated with P-Rex1, Rac1 and Cdc42, suggesting a scaffolding function for this pseudokinase. NRBP1-mediated promotion of cell migration and invasion was P-Rex1-dependent, while constitutively-active Rac1 rescued the effect of NRBP1 knockdown on cell proliferation and invasion. Generation of reactive oxygen species via a NRBP1/P-Rex1 pathway was implicated in these oncogenic roles of NRBP1. Overall, these findings define a new function for NRBP1 and a novel oncogenic signalling pathway in TNBC that may be amenable to therapeutic intervention.

show abstract

Section: Discussionmentioning

confidence: 81%

The pseudokinase NRBP1 activates Rac1/Cdc42 via P-Rex1 to drive oncogenic signalling in triple-negative breast cancer

et al. 2023

View full text Add to dashboard Cite

show abstract

“…SifA and DOCK180 bind ELMO1-PHD, they do so via two distinct and non-overlapping interfaces (Figure 3G). To experimentally validate this finding, we generated a mutant ELMO1 (W665A) that was previously confirmed by two independent groups to be essential for binding DOCK180 32,33 and tested its ability to bind His-SifA in pulldown assays. Both WT and ELMO1-W665A bound SifA to similar extents, indicating that W665 is dispensable for binding SifA (Figure 3H).…”

Section: Structure Homology Models Of Ternary Complexes Of Sifa•elmo1...mentioning

confidence: 94%

Diverse gut pathogens exploit the host engulfment pathway via a conserved mechanism

Anandachar

Agyekum

Sinha

et al. 2023

Preprint

View full text Add to dashboard Cite

Macrophages clear infections by engulfing and digesting pathogens within phagolysosomes. Pathogens escape this fate by engaging in a molecular arms race; they use WxxxE motif-containing effector proteins to subvert the host cells they invade and seek refuge within protective vacuoles. Here we define the host component of the molecular arms race as an evolutionarily conserved polar hotspot on the PH-domain of ELMO1 (Engulfment and Cell Motility1), which is targeted by diverse WxxxE-effectors. Using homology modeling and site-directed mutagenesis, we show that a lysine triad within the patch directly binds all WxxxE-effectors tested: SifA (Salmonella), IpgB1 and IpgB2 (Shigella), and Map (enteropathogenic E. coli). Using an integrated SifA-host protein-protein interaction (PPI) network, in-silico network perturbation, and functional studies we show that the major consequences of preventing SifA-ELMO1 interaction are reduced Rac1 activity and microbial invasion. That multiple effectors, all diverse in structure, function, and sequence bind the same hotpot on ELMO1 suggests that the WxxxE-effector(s)-ELMO1 interface is a convergence point of intrusion detection and/or host vulnerability. We conclude that the interface may represent the fault line in co-evolved molecular adaptations between pathogens and the host and its disruption may serve as a therapeutic strategy.

show abstract

“…RhoG activates Rac signaling during cell migration 6,7 and engulfment 10 via ELMO1/DOCK180 complex. The current model suggests the GTP-bound RhoG recruits the ELMO1 N-terminal RBD to the plasma membrane, while the C-terminal PH domain stabilizes binding of DOCK DHR-2 domain to Rac1 11,12 . This model of Rac1 activation through ELMO1/DOCK axis unveiled novel interfaces, such as PH/DHR-2 and ELMO1-RBD/RhoG, that are potentially explorable in drugging actin assembly.…”

mentioning

confidence: 84%

Targeting Ras-binding domain of ELMO1 by computational nanobody design

et al. 2023

Self Cite

View full text Add to dashboard Cite

The control of cell movement through manipulation of cytoskeletal structure has therapeutic prospects notably in the development of novel anti-metastatic drugs. In this study, we determine the structure of Ras-binding domain (RBD) of ELMO1, a protein involved in cytoskeletal regulation, both alone and in complex with the activator RhoG and verify its targetability through computational nanobody design. Using our dock-and-design approach optimized with native-like initial pose selection, we obtain Nb01, a detectable binder from scratch in the first-round design. An affinity maturation step guided by structure-activity relationship at the interface generates 23 Nb01 sequence variants and 17 of them show enhanced binding to ELMO1-RBD and are modeled to form major spatial overlaps with RhoG. The best binder, Nb29, inhibited ELMO1-RBD/RhoG interaction. Molecular dynamics simulation of the flexibility of CDR2 and CDR3 of Nb29 reveal the design of stabilizing mutations at the CDR-framework junctions potentially confers the affinity enhancement.

show abstract

Cryo-EM structure of the human ELMO1-DOCK5-Rac1 complex

Cited by 19 publications

References 64 publications

The pseudokinase NRBP1 activates Rac1/Cdc42 via P-Rex1 to drive oncogenic signalling in triple-negative breast cancer

The pseudokinase NRBP1 activates Rac1/Cdc42 via P-Rex1 to drive oncogenic signalling in triple-negative breast cancer

Diverse gut pathogens exploit the host engulfment pathway via a conserved mechanism

Targeting Ras-binding domain of ELMO1 by computational nanobody design

Contact Info

Product

Resources

About