Saka Hector Alex, Thompson J Will, Chen Yi-Shan, Dubois Laura G, Haas Joel T, Moseley Arthur, Valdivia Raphael H
Department of Molecular Genetics and Microbiology and Center for Microbial Pathogenesis, Duke University Medical Center, Durham, North Carolina, United States of America.
Proteomics and Metabolomics Core Facility, Duke University Medical Center, Durham, North Carolina, United States of America.
PLoS One. 2015 Apr 24;10(4):e0124630. doi: 10.1371/journal.pone.0124630. eCollection 2015.
The obligate intracellular bacterium Chlamydia trachomatis is a major human pathogen and a main cause of genital and ocular diseases. During its intracellular cycle, C. trachomatis replicates inside a membrane-bound vacuole termed an "inclusion". Acquisition of lipids (and other nutrients) from the host cell is a critical step in chlamydial replication. Lipid droplets (LD) are ubiquitous, ER-derived neutral lipid-rich storage organelles surrounded by a phospholipids monolayer and associated proteins. Previous studies have shown that LDs accumulate at the periphery of, and eventually translocate into, the chlamydial inclusion. These observations point out to Chlamydia-mediated manipulation of LDs in infected cells, which may impact the function and thereby the protein composition of these organelles. By means of a label-free quantitative mass spectrometry approach we found that the LD proteome is modified in the context of C. trachomatis infection. We determined that LDs isolated from C. trachomatis-infected cells were enriched in proteins related to lipid metabolism, biosynthesis and LD-specific functions. Interestingly, consistent with the observation that LDs intimately associate with the inclusion, a subset of inclusion membrane proteins co-purified with LD protein extracts. Finally, genetic ablation of LDs negatively affected generation of C. trachomatis infectious progeny, consistent with a role for LD biogenesis in optimal chlamydial growth.
专性胞内细菌沙眼衣原体是一种主要的人类病原体,也是引起生殖器和眼部疾病的主要原因。在其胞内周期中,沙眼衣原体在一种称为“包涵体”的膜结合空泡内进行复制。从宿主细胞获取脂质(和其他营养物质)是衣原体复制的关键步骤。脂滴(LD)广泛存在,是源自内质网的富含中性脂质的储存细胞器,被磷脂单层和相关蛋白所包围。先前的研究表明,脂滴在沙眼衣原体包涵体的周边积累,并最终转移到包涵体内。这些观察结果表明,衣原体在感染细胞中对脂滴进行了调控,这可能会影响这些细胞器的功能,进而影响其蛋白质组成。通过无标记定量质谱方法,我们发现脂滴蛋白质组在沙眼衣原体感染的情况下发生了改变。我们确定,从沙眼衣原体感染细胞中分离出的脂滴富含与脂质代谢、生物合成和脂滴特定功能相关的蛋白质。有趣的是,与脂滴与包涵体密切相关的观察结果一致,一部分包涵体膜蛋白与脂滴蛋白提取物共同纯化。最后,脂滴的基因消融对沙眼衣原体感染性后代的产生产生了负面影响,这与脂滴生物合成在衣原体最佳生长中的作用一致。