Crystallization and preliminary crystallographic analysis of the N-terminal actin binding domain of human fimbrin

Goldsmith, Sharon C.; Pokala, Navin; Matsudaira, Paul; Almo, Steven C.

doi:10.1002/(sici)1097-0134(199707)28:3<452::aid-prot13>3.0.co;2-g

Cited by 3 publications

(3 citation statements)

References 13 publications

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“…The requirement of SipA for efficient S. typhimurium internalization into nonphagocytic cells coupled to its ability to modulate the actin-bundling activity of T-plastin prompted us to investigate the role of T-plastin in Salmonella entry into cultured cells. COS-1 cells were transiently transfected with plasmids expressing a dominant negative form of T-plastin (43,44), and the ability of S. typhimurium to enter into those cells over time was examined by using a fluorescence microscopy assay (7). Expression of a dominant-negative form of T-plastin significantly reduced the levels of S. typhimurium internalization (Fig.…”

Section: Resultsmentioning

confidence: 63%

An invasion-associated Salmonella protein modulates the actin-bundling activity of plastin

Zhou

Mooseker

Galán

1999

Proc. Natl. Acad. Sci. U.S.A.

187

150

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The entry of Salmonella typhimurium into nonphagocytic cells requires a panel of bacterial effector proteins that are delivered to the host cell via a type III secretion system. These proteins modulate host-cell signal-transduction pathways and the actin cytoskeleton to induce membrane ruffling and bacterial internalization. One of these bacterial effectors, termed SipA, is an actin-binding protein that is required for efficient Salmonella entry into host cells. We report here that SipA forms a complex with T-plastin on bacterial infection. Formation of such a complex, which requires the presence of F-actin, results in a marked increase in the actin-bundling activity of T-plastin. We also report that T-plastin is recruited to S. typhimurium-induced membrane ruffles by a CDC42-dependent signaling process and is required for bacterial entry. We propose that modulation of the actin-bundling activity of T-plastin by SipA results in the stabilization of the actin filaments at the point of bacterial-host cell contact, which leads to more efficient Salmonella internalization.

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Section: Resultsmentioning

confidence: 63%

An invasion-associated Salmonella protein modulates the actin-bundling activity of plastin

Zhou

Mooseker

Galán

1999

Proc. Natl. Acad. Sci. U.S.A.

187

150

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“…The p.Glu270 and p.Trp499 residues affected in families 1 and 2, respectively, are contained in the core residues involved in actin binding (ABS, acting binding sites) 32,33 . Although the p.Glu270Lys variant (family 1) is not predicted to induce conformational changes, it does disrupt the bond between Gly270 and Lys236 (Fig 3B, magenta), which has been shown to be necessary for binding with F-actin 25 . The p.Trp499Cys change (family 2) in the ABD2 domain is predicted to affect the hydrophobic core of the actin-binding site (Fig 3B, yellow).…”

Section: Resultsmentioning

confidence: 99%

“…Structural modeling is based on the N-terminal actin-crosslinking domain structure from human plastin 3 obtained by X-ray diffraction(1AOA) 25 , on the actin-crosslinking core of Arabidopsis fimbrin (1PXY) by X-ray diffraction 26 , and on the fourth CH domain from human plastin 3 T-isoform (IWJC) by MNR (PDB ID: 1WJO) [unpublished data by deposition authors: Tomizawa, T., Kigawa, T., Koshiba, S., Inoue, M., Yokoyama, S., RIKEN Structural Genomics/Proteomics Initiative (RSGI)]. Data files can be downloaded from the Protein Data Bank (PDB, www.rcsb.org).…”

Section: Methodsmentioning

confidence: 99%

Missense variants affecting the actin-binding domains of PLS3 cause X-linked congenital diaphragmatic hernia and body wall defects

Petit

Longoni

Wells

et al. 2021

Preprint

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Congenital diaphragmatic hernia (CDH) is a relatively common and genetically heterogeneous structural birth defect associated with high mortality and morbidity. We describe eight unrelated families with a novel X-linked condition characterized by diaphragm defects, variable anterior body wall anomalies, and/or facial dysmorphism. Using linkage analysis and whole exome or whole genome sequencing, we identified novel missense variants in the actin binding domains of plastin 3 (PLS3), a gene encoding an actin bundling protein, that co-segregate with disease in all families. Loss-of-function variants in PLS3 have been described previously in association with X-linked osteoporosis. To address these seemingly disparate clinical phenotypes, we performed in silico protein modeling and cellular overexpression experiments, which suggest that the affected residues in individuals with CDH are important for actin binding and result in disorganization of the actin cytoskeleton and a reduction in normal actin stress fiber formation. A mouse knock-in model of a variant identified in one of the families, p.W499C, shows partial perinatal lethality and recapitulates the key findings of the human phenotype, including diaphragm and abdominal wall defects. Both the mouse model and one surviving adult patient with a PLS3 variant were observed to have increased, rather than decreased, bone mineral density. Together, these clinical and functional data in human and mouse reveal that specific missense variants affecting the actin binding domains of PLS3 may have a gain-of-function effect and cause a new Mendelian disorder.

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PLS3 missense variants affecting the actin-binding domains cause X-linked congenital diaphragmatic hernia and body-wall defects

Petit,

Longoni,

Wells

et al. 2023

The American Journal of Human Genetics

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Crystallization and preliminary crystallographic analysis of the N-terminal actin binding domain of human fimbrin

Cited by 3 publications

References 13 publications

An invasion-associated Salmonella protein modulates the actin-bundling activity of plastin

An invasion-associated Salmonella protein modulates the actin-bundling activity of plastin

Missense variants affecting the actin-binding domains of PLS3 cause X-linked congenital diaphragmatic hernia and body wall defects

PLS3 missense variants affecting the actin-binding domains cause X-linked congenital diaphragmatic hernia and body-wall defects

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