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Protochlamydia Induces Apoptosis of Human HEp-2 Cells through Mitochondrial Dysfunction Mediated by Chlamydial Protease-Like Activity Factor
Obligate amoebal endosymbiotic bacterium Protochlamydia with ancestral pathogenic chlamydial features evolved to survive within protist hosts, such as Acanthamoba, 0.7–1.4 billion years ago, but not within vertebrates including humans. This observation raises the possibility that interactions betwee...
Autores principales: | , , , , , , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Public Library of Science
2013
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3569409/ https://www.ncbi.nlm.nih.gov/pubmed/23409113 http://dx.doi.org/10.1371/journal.pone.0056005 |
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author | Matsuo, Junji Nakamura, Shinji Ito, Atsushi Yamazaki, Tomohiro Ishida, Kasumi Hayashi, Yasuhiro Yoshida, Mitsutaka Takahashi, Kaori Sekizuka, Tsuyoshi Takeuchi, Fumihiko Kuroda, Makoto Nagai, Hiroki Hayashida, Kyoko Sugimoto, Chihiro Yamaguchi, Hiroyuki |
author_facet | Matsuo, Junji Nakamura, Shinji Ito, Atsushi Yamazaki, Tomohiro Ishida, Kasumi Hayashi, Yasuhiro Yoshida, Mitsutaka Takahashi, Kaori Sekizuka, Tsuyoshi Takeuchi, Fumihiko Kuroda, Makoto Nagai, Hiroki Hayashida, Kyoko Sugimoto, Chihiro Yamaguchi, Hiroyuki |
author_sort | Matsuo, Junji |
collection | PubMed |
description | Obligate amoebal endosymbiotic bacterium Protochlamydia with ancestral pathogenic chlamydial features evolved to survive within protist hosts, such as Acanthamoba, 0.7–1.4 billion years ago, but not within vertebrates including humans. This observation raises the possibility that interactions between Protochlamydia and human cells may result in a novel cytopathic effect, leading to new insights into host-parasite relationships. Previously, we reported that Protochlamydia induces apoptosis of the immortalized human cell line, HEp-2. In this study, we attempted to elucidate the molecular mechanism underlying this apoptosis. We first confirmed that, upon stimulation with the bacteria, poly (ADP-ribose) polymerase (PARP) was cleaved at an early stage in HEp-2 cells, which was dependent on the amount of bacteria. A pan-caspase inhibitor and both caspase-3 and -9 inhibitors similarly inhibited the apoptosis of HEp-2 cells. A decrease of the mitochondrial membrane potential was also confirmed. Furthermore, lactacystin, an inhibitor of chlamydial protease-like activity factor (CPAF), blocked the apoptosis. Cytochalasin D also inhibited the apoptosis, which was dependent on the drug concentration, indicating that bacterial entry into cells was required to induce apoptosis. Interestingly, Yersinia type III inhibitors (ME0052, ME0053, and ME0054) did not have any effect on the apoptosis. We also confirmed that the Protochlamydia used in this study possessed a homologue of the cpaf gene and that two critical residues, histidine-101 and serine-499 of C. trachomatis CPAF in the active center, were conserved. Thus, our results indicate that after entry, Protochlamydia-secreted CPAF induces mitochondrial dysfunction with a decrease of the membrane potential, followed by caspase-9, caspase-3 and PARP cleavages for apoptosis. More interestingly, because C. trachomatis infection can block the apoptosis, our finding implies unique features of CPAF between pathogenic and primitive chlamydiae. |
format | Online Article Text |
id | pubmed-3569409 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2013 |
publisher | Public Library of Science |
record_format | MEDLINE/PubMed |
spelling | pubmed-35694092013-02-13 Protochlamydia Induces Apoptosis of Human HEp-2 Cells through Mitochondrial Dysfunction Mediated by Chlamydial Protease-Like Activity Factor Matsuo, Junji Nakamura, Shinji Ito, Atsushi Yamazaki, Tomohiro Ishida, Kasumi Hayashi, Yasuhiro Yoshida, Mitsutaka Takahashi, Kaori Sekizuka, Tsuyoshi Takeuchi, Fumihiko Kuroda, Makoto Nagai, Hiroki Hayashida, Kyoko Sugimoto, Chihiro Yamaguchi, Hiroyuki PLoS One Research Article Obligate amoebal endosymbiotic bacterium Protochlamydia with ancestral pathogenic chlamydial features evolved to survive within protist hosts, such as Acanthamoba, 0.7–1.4 billion years ago, but not within vertebrates including humans. This observation raises the possibility that interactions between Protochlamydia and human cells may result in a novel cytopathic effect, leading to new insights into host-parasite relationships. Previously, we reported that Protochlamydia induces apoptosis of the immortalized human cell line, HEp-2. In this study, we attempted to elucidate the molecular mechanism underlying this apoptosis. We first confirmed that, upon stimulation with the bacteria, poly (ADP-ribose) polymerase (PARP) was cleaved at an early stage in HEp-2 cells, which was dependent on the amount of bacteria. A pan-caspase inhibitor and both caspase-3 and -9 inhibitors similarly inhibited the apoptosis of HEp-2 cells. A decrease of the mitochondrial membrane potential was also confirmed. Furthermore, lactacystin, an inhibitor of chlamydial protease-like activity factor (CPAF), blocked the apoptosis. Cytochalasin D also inhibited the apoptosis, which was dependent on the drug concentration, indicating that bacterial entry into cells was required to induce apoptosis. Interestingly, Yersinia type III inhibitors (ME0052, ME0053, and ME0054) did not have any effect on the apoptosis. We also confirmed that the Protochlamydia used in this study possessed a homologue of the cpaf gene and that two critical residues, histidine-101 and serine-499 of C. trachomatis CPAF in the active center, were conserved. Thus, our results indicate that after entry, Protochlamydia-secreted CPAF induces mitochondrial dysfunction with a decrease of the membrane potential, followed by caspase-9, caspase-3 and PARP cleavages for apoptosis. More interestingly, because C. trachomatis infection can block the apoptosis, our finding implies unique features of CPAF between pathogenic and primitive chlamydiae. Public Library of Science 2013-02-11 /pmc/articles/PMC3569409/ /pubmed/23409113 http://dx.doi.org/10.1371/journal.pone.0056005 Text en © 2013 Matsuo et al http://creativecommons.org/licenses/by/4.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are properly credited. |
spellingShingle | Research Article Matsuo, Junji Nakamura, Shinji Ito, Atsushi Yamazaki, Tomohiro Ishida, Kasumi Hayashi, Yasuhiro Yoshida, Mitsutaka Takahashi, Kaori Sekizuka, Tsuyoshi Takeuchi, Fumihiko Kuroda, Makoto Nagai, Hiroki Hayashida, Kyoko Sugimoto, Chihiro Yamaguchi, Hiroyuki Protochlamydia Induces Apoptosis of Human HEp-2 Cells through Mitochondrial Dysfunction Mediated by Chlamydial Protease-Like Activity Factor |
title |
Protochlamydia Induces Apoptosis of Human HEp-2 Cells through Mitochondrial Dysfunction Mediated by Chlamydial Protease-Like Activity Factor |
title_full |
Protochlamydia Induces Apoptosis of Human HEp-2 Cells through Mitochondrial Dysfunction Mediated by Chlamydial Protease-Like Activity Factor |
title_fullStr |
Protochlamydia Induces Apoptosis of Human HEp-2 Cells through Mitochondrial Dysfunction Mediated by Chlamydial Protease-Like Activity Factor |
title_full_unstemmed |
Protochlamydia Induces Apoptosis of Human HEp-2 Cells through Mitochondrial Dysfunction Mediated by Chlamydial Protease-Like Activity Factor |
title_short |
Protochlamydia Induces Apoptosis of Human HEp-2 Cells through Mitochondrial Dysfunction Mediated by Chlamydial Protease-Like Activity Factor |
title_sort | protochlamydia induces apoptosis of human hep-2 cells through mitochondrial dysfunction mediated by chlamydial protease-like activity factor |
topic | Research Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3569409/ https://www.ncbi.nlm.nih.gov/pubmed/23409113 http://dx.doi.org/10.1371/journal.pone.0056005 |
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