Autoinhibition and activation mechanisms of the Wiskott–Aldrich syndrome protein

Kim, Annette S.; Kakalis, Lazaros T.; Abdul-Manan, Norzehan; Liu, Grace A.; Rosen, Michael K.

doi:10.1038/35004513

Cited by 656 publications

(733 citation statements)

References 43 publications

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“…The WASP C-terminal VCA domain interacts with Arp2/3 complex and stimulates actin polymerization, thereby resulting in formation of filopodia (7,8). This WASP C-terminal activity is mainly regulated by Cdc42.…”

Section: The Role Of the Wasp-wip Complex In Podosome Formationmentioning

confidence: 99%

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Requirement for a Complex of Wiskott-Aldrich Syndrome Protein (WASP) with WASP Interacting Protein in Podosome Formation in Macrophages

Tsuboi

2007

The Journal of Immunology

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Chemotactic migration of macrophages is critical for the recruitment of leukocytes to inflamed tissues. Macrophages use a specialized adhesive structure called a podosome to migrate. Podosome formation requires the Wiskott-Aldrich syndrome protein (WASP), which is a product of the gene defective in an X-linked inherited immunodeficiency disorder, the Wiskott-Aldrich syndrome. Macrophages from WASP-deficient Wiskott-Aldrich syndrome patients lack podosomes, resulting in defective chemotactic migration. However, the molecular basis for podosome formation is not fully understood. I have shown that the WASP interacting protein (WIP), a binding partner of WASP, plays an important role in podosome formation in macrophages. I showed that WASP bound WIP to form a complex at podosomes and that the knockdown of WIP impairs podosome formation. When WASP binding to WIP was blocked, podosome formation was also impaired. When WASP expression was reduced by small interfering RNA transfection, the amount of the complex of WASP with WIP decreased, resulting in reduced podosome formation. Podosomes were restored by reconstitution of the WASP-WIP complex in WASP knockdown cells. These results indicate that the WASP-WIP complex is required for podosome formation in macrophages. When podosome formation was reduced by blocking WASP binding to WIP, transendothelial migration of macrophages, the most crucial process in macrophage trafficking, was impaired. These results suggest that a complex of WASP with WIP plays a critical role in podosome formation, thereby mediating efficient transendothelial migration of macrophages.

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Section: The Role Of the Wasp-wip Complex In Podosome Formationmentioning

confidence: 99%

“…N-WASP, the more widely expressed homologue, has the same domain organization as WASP (6). The WASP C-terminal verprolin/cofilin/acidic (VCA) domain stimulates actin polymerization by interacting with an actin-related protein complex, the actin-related protein 2/3 (Arp2/3) complex (7,8).…”

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confidence: 99%

Requirement for a Complex of Wiskott-Aldrich Syndrome Protein (WASP) with WASP Interacting Protein in Podosome Formation in Macrophages

Tsuboi

2007

The Journal of Immunology

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“…The plasticity afforded by the presence of a flexible structure with multiple conformations can allow a single protein to recognize a number of biological targets (38), and mediate different binding events (37). Lack of structure in the free state may also result in an increase in binding specificity, since the energetic cost of structural rearrangement must be compensated by a higher binding energy (39).…”

Section: Both Lc8 and Tctex-1 Bind To The N-terminal Domain Of Ic74mentioning

confidence: 99%

Interactions of Cytoplasmic Dynein Light Chains Tctex-1 and LC8 with the Intermediate Chain IC74

et al. 2002

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The interactions of three subunits of cytoplasmic dynein from Drosophila melanogaster, LC8, Tctex-1, and the N-terminal domain of IC74 (N-IC74, residues 1-289), were characterized in vitro by affinity methods, limited proteolysis, and circular dichroism spectroscopy. These subunits were chosen for study because they are presumed to promote the assembly of the complex and to be engaged in the controlled binding and release of cargo. Limited proteolysis and mass spectrometry of N-IC74 in the presence of LC8 and Tctex-1 localized binding of Tctex-1 to the vicinity of K104 and K105, and localized binding of LC8 to the region downstream of K130. Circular dichroism, fluorescence, sedimentation velocity, and proteolysis studies indicate that N-IC74 has limited secondary and tertiary structure at near physiological solution conditions. Upon addition of LC8, N-IC74 undergoes a significant conformational change from largely unfolded to a more ordered structure. This conformational change is reflected in increased global protection of N-IC74 from proteolytic digestion following the interaction, and in a significant change in the CD signal. A smaller but reproducible change in the CD spectra was observed upon Tctex-1 binding as well. The increased structure introduced into N-IC74 upon light chain binding suggests a mechanism by which LC8 and Tctex-1 may regulate the assembly of the dynein complex.

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“…The multiple domains integrate signals that induce the following: 1) WASP recruitment to activation sites (e.g., immune synapse); and 2) conformational activation by exposure of the Cterminal VCA (verprolin/cofilin/acidic) domains that catalyze nucleation of actin filaments (8,9). WASP is found in the cytoplasm of resting cells in the inactive autoinhibited conformation in which the VCA domains are bound to the central GTPase binding domain (GBD) (10). The resting cell molecule is further stabilized by binding of WIP to the large N-terminal Ena/VASP homology 1 (EVH1) domain (11,12).…”

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Genotype-Proteotype Linkage in the Wiskott-Aldrich Syndrome

Lutskiy

Rosen

Remold‐O′Donnell

2005

The Journal of Immunology

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Wiskott-Aldrich syndrome (WAS) is a platelet/immunodeficiency disease arising from mutations of WAS protein (WASP), a hemopoietic cytoskeletal protein. Clinical symptoms vary widely from mild (X-linked thrombocytopenia) to life threatening. In this study, we examined the molecular effects of individual mutations by quantifying WASP in peripheral lymphocytes of 44 patients and identifying the molecular variant (collectively called proteotype). Nonpredicted proteotypes were found for 14 genotypes. These include WASP-negative lymphocytes found for five missense genotypes and WASP-positive lymphocytes for two nonsense, five frameshift, and two splice site genotypes. Missense mutations in the Ena/VASP homology 1 (EVH1) domain lead to decreased/absent WASP but normal mRNA levels, indicating that proteolysis causes the protein deficit. Because several of the EVH1 missense mutations alter WIP binding sites, the findings suggest that abrogation of WIP binding induces proteolysis. Whereas platelets of most patients were previously shown to lack WASP, WASP-positive platelets were found for two atypical patients, both of whom have mutations outside the EVH1 domain. WASP variants with alternative splicing and intact C-terminal domains were characterized for eight nonsense and frameshift genotypes. One of these, a nonsense genotype in a mild patient, supports expression of WASP lacking half of the proline-rich region. With one notable exception, genotype and proteotype were linked, indicating that a genotype-proteotype registry could be assembled to aid in predicting disease course and planning therapy for newly diagnosed infants. Knowledge of the molecular effect of mutations would aid also in identifying disease-modifying genes.

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Autoinhibition and activation mechanisms of the Wiskott–Aldrich syndrome protein

Cited by 656 publications

References 43 publications

Requirement for a Complex of Wiskott-Aldrich Syndrome Protein (WASP) with WASP Interacting Protein in Podosome Formation in Macrophages

Requirement for a Complex of Wiskott-Aldrich Syndrome Protein (WASP) with WASP Interacting Protein in Podosome Formation in Macrophages

Interactions of Cytoplasmic Dynein Light Chains Tctex-1 and LC8 with the Intermediate Chain IC74

Genotype-Proteotype Linkage in the Wiskott-Aldrich Syndrome

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