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Cited by 173Protein Kinase Regulation and GTPase SignalingCellular Mechanics and InteractionsCellular transport and secretion

Rac1 GTPase activates the WAVE regulatory complex through two distinct binding sites

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Chen, Baoyu, Chou, Hui-Ting, Brautigam, Chad A, Xing, Wenmin, Yang, Sheng, Henry, Lisa, Doolittle, Lynda K, Walz, Thomas +1 more·2017·eLife
DOI 10.7554/elife.29795PMC PMC5614565
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Abstract

The Rho GTPase Rac1 activates the WAVE regulatory complex (WRC) to drive Arp2/3 complex-mediated actin polymerization, which underpins diverse cellular processes. Here we report the structure of a WRC-Rac1 complex determined by cryo-electron microscopy. Surprisingly, Rac1 is not located at the binding site on the Sra1 subunit of the WRC previously identified by mutagenesis and biochemical data. Rather, it binds to a distinct, conserved site on the opposite end of Sra1. Biophysical and biochemical data on WRC mutants confirm that Rac1 binds to both sites, with the newly identified site having higher affinity and both sites required for WRC activation. Our data reveal that the WRC is activated by simultaneous engagement of two Rac1 molecules, suggesting a mechanism by which cells may sense the density of active Rac1 at membranes to precisely control actin assembly.

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Crossref · 10.7554/elife.29795

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  • Cryo-electron microscopy of the WAVE complex with RAC1 indicated that the Ser968Phe mutation in CYFIP1 (corresponding to Ser968Phe in CYFIP2) is located just beneath the RAC1 binding site.

    “Alternatively, cryo-electron microscopy of the WAVE complex with RAC1 indicated that the Ser968Phe mutation in CYFIP1 (corresponding to Ser968Phe in CYFIP2) is located just beneath the RAC1 binding site.”

    From The WAVE complex in developmental and adulthood brain disorders (2025) · cites this paper as [172]

    The Results and Discussion place S969F in Sra1 (CYFIP1) immediately beneath the D Site, a Rac1-binding site, and identify S968F in CYFIP2 as the equivalent mutation. The claim gets the location right but gives the CYFIP1 residue as Ser968 rather than the paper’s Ser969. (LLM, conf=0.90)

    Experimental — accuracy not yet measured

  • Ser968Phe in CYFIP1 facilitates RAC1 binding and WAVE complex activation.

    “Further characterization revealed that Ser968Phe in CYFIP1 facilitates RAC1 binding and WAVE complex activation172.”

    From The WAVE complex in developmental and adulthood brain disorders (2025) · cites this paper as [172]

    The Discussion (page unavailable) reports that S969F in Sra1 facilitates Rac1 binding and WAVE regulatory complex activation. It identifies S968F as the corresponding mutation in CYFIP2, not CYFIP1, so the citing sentence misidentifies the mutation studied. (LLM, conf=0.90)

    Experimental — accuracy not yet measured

  • Two distinct RAC1-binding sites on CYFIP1/2 and a recently discovered ARF1-binding site on HEM1/2 cooperatively activate the WRC.

    “Two distinct RAC1-binding sites on CYFIP1/2 and a recently discovered ARF1-binding site on HEM1/2 cooperatively activate the WRC [16, 17].”

    From HEM1 Actin Immunodysregulatory Disorder: Genotypes, Phenotypes, and Future Directions (2022) · cites this paper as [17]

    The Introduction and Discussion report two Rac1-binding sites on Sra1 and show that both contribute to WRC activation. The Discussion mentions prior observations that Rac1 and Arf can cooperatively activate the WRC, but it does not locate an Arf1-binding site on HEM1/2 or establish cooperation among all three sites. (LLM, conf=0.88)

    Experimental — accuracy not yet measured

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Cited by

Sorted by citations

Authors
Chen, B., Chou, H.-T., Brautigam, C. A., Xing, W., Yang, S., Henry, L., Doolittle, L. K., Walz, T., & Rosen, M. K.
Affiliations
  • Howard Hughes Medical Institute (US)
  • The University of Texas Southwestern Medical Center (US)
  • Iowa State University (US)
  • Rockefeller University (US)
Corresponding author
Rosen, Michael K
ORCID
Venue
eLife
Publisher
eLife Sciences Publications, Ltd
Published
2017-09-26
Volume
6
ISSN
2050-084X (ISSN-L)
DOI
10.7554/elife.29795
PMC
PMC5614565
OpenAlex
W2759963226
Cited by
173
References
1
Citations by year
2026: 17 · 2025: 20 · 2024: 15 · 2023: 19 · 2022: 17 · 2021: 26 · 2020: 28 · 2019: 19 · 2018: 12
FWCI
6.42
Type
journal article
Language
en
Open access
Licence
Locations
  • eLife (publishedVersion, cc-by)
  • Europe PMC (PubMed Central) (publishedVersion, cc-by)
  • PubMed Central (publishedVersion, cc-by)
  • PubMed (publishedVersion)
  • Iowa State University Digital Repository (Iowa State University) (submittedVersion, cc-by)
  • Digital Access to Scholarship at Harvard (DASH) (Harvard University) (submittedVersion, cc-by)
  • Iowa State University Digital Repository (Iowa State University) (submittedVersion)
  • DOAJ (DOAJ: Directory of Open Access Journals) (submittedVersion, cc-by-sa)
Funders
  • National Institutes of Health (R01-056322)
  • Welch Foundation (I-1544)
  • Howard Hughes Medical Institute
  • Iowa State University & Roy J. Carver Charitable Trust
Topics
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Protein Kinase Regulation and GTPase Signaling
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Cell Biology › Cellular Mechanics and Interactions
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Cell Biology › Cellular transport and secretion
Keywords
Rac1, WAVE regulatory complex, SR-A1, Arp2/3 complex, actin polymerization, Rho GTPases, protein complex structure
MeSH terms
Actins, Binding Sites, Carrier Proteins, Cytoskeletal Proteins, DNA Mutational Analysis, Humans, Protein Binding, Proteins, tRNA Methyltransferases, Cryoelectron Microscopy, rac1 GTP-Binding Protein, Adaptor Proteins, Signal Transducing, Wiskott-Aldrich Syndrome Protein Family, Protein Multimerization

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