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Proc Natl Acad Sci U S A. 2013 Oct 15;110(42):E4007-15. doi: 10.1073/pnas.1316852110. Epub 2013 Oct 1.

Calpain-dependent cytoskeletal rearrangement exploited for anthrax toxin endocytosis.

Author information

1
Departments of Genetics and Medicine, Stanford University School of Medicine, Stanford, CA 94305.

Abstract

The protective antigen component of Bacillus anthracis toxins can interact with at least three distinct proteins on the host cell surface, capillary morphogenesis gene 2 (CMG2), tumor endothelial marker 8, and β1-integrin, and, with the assistance of other host proteins, enters targeted cells by receptor-mediated endocytosis. Using an antisense-based phenotypic screen, we discovered the role of calpains in this process. We show that functions of a ubiquitous Ca(2+)-dependent cysteine protease, calpain-2, and of the calpain substrate talin-1 are exploited for association of anthrax toxin and its principal receptor, CMG2, with higher-order actin filaments and consequently for toxin entry into host cells. Down-regulated expression of calpain-2 or talin-1, or pharmacological interference with calpain action, did not affect toxin binding but reduced endocytosis and increased the survival of cells exposed to anthrax lethal toxin. Adventitious expression of wild-type talin-1 promoted toxin endocytosis and lethality, whereas expression of a talin-1 mutant (L432G) that is insensitive to calpain cleavage did not. Disruption of talin-1, which links integrin-containing focal adhesion complexes to the actin cytoskeleton, facilitated association of toxin bound to its principal cell-surface receptor, CMG2, with higher-order actin filaments undergoing dynamic disassembly and reassembly during endocytosis. Our results reveal a mechanism by which a bacterial toxin uses constitutively occurring calpain-mediated cytoskeletal rearrangement for internalization.

KEYWORDS:

MDL28170; calpastatin; lethal factor

PMID:
24085852
PMCID:
PMC3801034
DOI:
10.1073/pnas.1316852110
[Indexed for MEDLINE]
Free PMC Article

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