Cdc42 and the Actin-Related Protein/Neural Wiskott-Aldrich Syndrome Protein Network Mediate Cellular Invasion by Cryptosporidium parvum

Xian Ming Chen, Bing Q. Huang, Patrick L. Splinter, James D. Orth, Daniel D Billadeau, Mark A Mc Niven, Nicholas F La Russo

Research output: Contribution to journalArticle

38 Citations (Scopus)

Abstract

Cryptosporidium parvum invasion of epithelial cells involves host cell membrane alterations which require a remodeling of the host cell actin cytoskeleton. In addition, an actin plaque, possibly associated with the dense-band region, forms within the host cytoplasm at the host-parasite interface. Here we show that Cdc42 and RhoA, but not Rac1, members of the Rho family of GTPases, are recruited to the host-parasite interface in an in vitro model of human biliary cryptosporidiosis. Interestingly, activation of Cdc42, but not RhoA, was detected in the infected cells. Neural Wiskott-Aldrich syndrome protein (N-WASP) and p34-Arc, actin-regulating downstream effectors of Cdc42, were also recruited to the host-parasite interface. Whereas cellular expression of a constitutively active mutant of Cdc42 promoted C. parvum invasion, overexpression of a dominant negative mutant of Cdc42, or depletion of Cdc42 mRNA by short interfering RNA-mediated gene silencing, inhibited C. parvum invasion. Expression of the WA fragment of N-WASP to block associated actin polymerization also inhibited C. parvum invasion. Moreover, inhibition of host cell Cdc42 activation by dominant negative mutation inhibited C. parvum-associated actin remodeling, membrane protrusion, and dense-band formation. In contrast, treatment of cells with a Rho inhibitor, exoenzyme C3, or cellular overexpression of dominant negative mutants of RhoA and Rac1 had no effect on C. parvum invasion. These data suggest that C. parvum invasion of target epithelia results from the organism's ability to activate a host cell Cdc42 GTPase signaling pathway to induce host cell actin remodeling at the attachment site.

Original languageEnglish (US)
Pages (from-to)3011-3021
Number of pages11
JournalInfection and Immunity
Volume72
Issue number5
DOIs
StatePublished - May 2004

Fingerprint

Wiskott-Aldrich Syndrome Protein
Cryptosporidium parvum
Actins
Proteins
Parasites
Cryptosporidiosis
rho GTP-Binding Proteins
Aptitude
GTP Phosphohydrolases
Gene Silencing
Actin Cytoskeleton
Polymerization
Small Interfering RNA
Cytoplasm
Epithelium
Epithelial Cells
Cell Membrane
Messenger RNA
Mutation
Membranes

ASJC Scopus subject areas

  • Immunology

Cite this

Cdc42 and the Actin-Related Protein/Neural Wiskott-Aldrich Syndrome Protein Network Mediate Cellular Invasion by Cryptosporidium parvum. / Chen, Xian Ming; Huang, Bing Q.; Splinter, Patrick L.; Orth, James D.; Billadeau, Daniel D; Mc Niven, Mark A; La Russo, Nicholas F.

In: Infection and Immunity, Vol. 72, No. 5, 05.2004, p. 3011-3021.

Research output: Contribution to journalArticle

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