RRC ID 45420
著者 Ishida K, Matsuo J, Yamamoto Y, Yamaguchi H.
タイトル Chlamydia pneumoniae effector chlamydial outer protein N sequesters fructose bisphosphate aldolase A, providing a benefit to bacterial growth.
ジャーナル BMC Microbiol
Abstract BACKGROUND:Pathogenic chlamydiae are obligate intracellular pathogens and have adapted successfully to human cells, causing sexually transmitted diseases or pneumonia. Chlamydial outer protein N (CopN) is likely a critical effector protein secreted by the type III secretion system in chlamydiae, which manipulates host cells. However, the mechanisms of its action remain to be clarified. In this work, we aimed to identify previously unidentified CopN effector target in host cells.
RESULTS:We first performed a pull-down assay with recombinant glutathione S-transferase (GST) fusion CopN proteins (GST-CpCopN: Chlamydia pneumoniae TW183, GST-CtCopN: Chlamydia trachomatis D/UW-3/CX) as "bait" and soluble lysates obtained from human immortal epithelial HEp-2 cells as "prey", followed by SDS-PAGE with mass spectroscopy (MS). We found that a host cell protein specifically bound to GST-CpCopN, but not GST-CtCopN. MS revealed the host protein to be fructose bisphosphate aldolase A (aldolase A), which plays a key role in glycolytic metabolism. We also confirmed the role of aldolase A in chlamydia-infected HEp-2 cells by using two distinct experiments for gene knockdown with an siRNA specific to aldolase A transcripts, and for assessment of glycolytic enzyme gene expression levels. As a result, both the numbers of chlamydial inclusion-forming units and RpoD transcripts were increased in the chlamydia-infected aldolase A knockdown cells, as compared with the wild-type HEp-2 cells. Meanwhile, chlamydial infection tended to enhance expression of aldolase A.
CONCLUSIONS:We discovered that one of the C. pneumoniae CopN targets is the glycolytic enzyme aldolase A. Sequestering aldolase A may be beneficial to bacterial growth in infected host cells.
巻・号 14
ページ 330
公開日 2014-12-21
DOI 10.1186/s12866-014-0330-3
PII s12866-014-0330-3
PMID 25528659
PMC PMC4302594
MeSH Bacterial Outer Membrane Proteins / metabolism Cell Line Chlamydia Infections / metabolism Chlamydia trachomatis / metabolism Chlamydophila pneumoniae / metabolism* Fructose-Bisphosphate Aldolase / metabolism* Glycolysis / physiology Humans
IF 2.989
引用数 4
WOS 分野 MICROBIOLOGY
リソース情報
ヒト・動物細胞