The protein SdhA maintains the integrity of the Legionella-containing vacuole
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA
Authors: Creasey, Elizabeth A.; Isberg, Ralph R.
Abstract
Legionella pneumophila directs the formation of a specialized vacuole within host cells, dependent on protein substrates of the Icm/Dot translocation system. Survival of the host cell is essential for intracellular replication of L. pneumophila. Strains lacking the translocated substrate SdhA are defective for intracellular replication and activate host cell death pathways in primary macrophages. To understand how SdhA promotes evasion of death pathways, we performed a mutant hunt to identify bacterial suppressors of the Delta sdhA growth defect. We identified the secreted phospholipase PlaA as key to activation of death pathways by the Delta sdhA strain. Based on homology between PlaA and SseJ, a Salmonella protein associated with vacuole degradation, we determined the roles of SdhA and PlaA in controlling vacuole integrity. In the absence of sdhA, the Legionella-containing vacuole was unstable, resulting in access to the host cytosol. Both vacuole disruption and host cell death were largely dependent on PlaA. Consistent with these observations, the Delta sdhA strain colocalized with galectin - 3, a marker of vacuole rupture, in a PlaA - dependent process. Access of Delta sdhA strains to the macrophage cytosol triggered multiple responses in the host cell, including degradation of bacteria, induction of the type I IFN response, and activation of inflammasomes. Therefore, we have demonstrated that the Legionella - containing vacuole is actively stabilized by the SdhA protein during intracellular replication. This vacuolar niche affords the bacterium protection from cytosolic host factors that degrade bacteria and initiate immune responses.
Culture-independent analysis of the bacterial community during fermentation of pa-som, a traditional fermented fish product in Laos
FISHERIES SCIENCE
Authors: Marui, Junichiro; Boulom, Sayvisene; Panthavee, Wanchai; Momma, Mari; Kusumoto, Ken-Ichi; Nakahara, Kazuhiko; Saito, Masayoshi
Abstract
Pa-som is a traditional salty fermented food made from varieties of freshwater fish with garlic and rice in Laos. A similar product, plaa-som, is made in Thailand. To investigate the fermentation process of pa-som, lactic acid production and pH were monitored during laboratory-scale production of pa-som. Furthermore, the variety of bacteria involved and their succession during fermentation were examined using culture-independent polymerase chain reaction (PCR) denaturing gel gradient electrophoresis targeting the 16S ribosomal RNA (rRNA) gene. d- and l-lactic acid production and a decrease in pH were observed in the samples after 1 day of fermentation. The lactic acid content and pH continued to increase and decrease, respectively, until 4 days of fermentation. Overall, six lactic acid bacteria and eight other indigenous bacteria species were detected during pa-som fermentation. Among the lactic acid bacteria detected, two Lactococcus and one Weissella species were detected in samples after 1-4 days of fermentation, suggesting that these three species play major roles in pa-som fermentation from the initial phase. This is the first report to investigate the fermentation process in pa-som at the microbial level. This research approach should be applicable to a wide variety of pa-som and related fermented fishery products in Laos to help understand their microbial diversity and to identify beneficial bacterial species for improving quality.