title: Porphyromonas gingivalis influences actin degradation within epithelial cells during invasion and apoptosis.
abstract: Porphyromonas gingivalis, a Gram-negative oral pathogen, has been shown to induce apoptosis in human gingival epithelial cells, yet the underlining cellular mechanisms controlling this process are poorly understood. We have previously shown that the P. gingivalis proteases arginine and lysine gingipains, are necessary and sufficient to induce host cell apoptosis. In the present study, we demonstrate that 'P. gingivalis-induced apoptosis' is mediated through degradation of actin leading to cytoskeleton collapse. Stimulation of human gingival epithelial cells with P. gingivalis strains 33277 and W50 at moi:100 induced β-actin cleavage as early as 1 h and human serum inhibited this effect. By using gingipain-deficient mutants of P. gingivalis and purified gingipains, we demonstrate that lysine gingipain is involved in actin hydrolysis in a dose and time-dependent manner. Use of Jasplakinolide and cytochalasin D revealed that P. gingivalis internalization is necessary for actin cleavage. Further, we also show that lysine gingipain from P. gingivalis can cleave active caspase 3. Taken together, we have identified actin as a substrate for lysine gingipain and demonstrated a novel mechanism involved in microbial host cell invasion and apoptosis.
authors.affiliation: Department of Periodontics, School of Dental Medicine, University of Pennsylvania, Philadelphia, PA 19104, USA.
authors.forename: James A
authors.initials: JA
authors.lastname: Kinane
authors.forename: Manjunatha R
authors.initials: MR
authors.lastname: Benakanakere
authors.forename: Jiawei
authors.initials: J
authors.lastname: Zhao
authors.forename: Kavita B
authors.initials: KB
authors.lastname: Hosur
authors.forename: Denis F
authors.initials: DF
authors.lastname: Kinane
mesh_terms: D000199:Actins / Q000378:metabolism*; D018829:Adhesins, Bacterial / Q000378:metabolism; D017209:Apoptosis; D002478:Cells, Cultured; D003546:Cysteine Endopeptidases / Q000378:metabolism; D004847:Epithelial Cells / Q000382:microbiology*; D000080867:Gingipain Cysteine Endopeptidases; D006801:Humans; D016966:Porphyromonas gingivalis / Q000201:enzymology / Q000472:pathogenicity*
chemical_list: D000199:Actins; D018829:Adhesins, Bacterial; D000080867:Gingipain Cysteine Endopeptidases; D003546:Cysteine Endopeptidases