Ma Liang, Jia Xiao-Hui, Gao Zhe, Zhou Yan, Cheng Yong-Ting, Li Ping, Jia Tian-Jun
Pathogen Biology and Immunology Research Institute, Hebei North University, Zhangjiakou, Hebei, China.
Key Laboratory of Clinical Laboratory Diagnostics, Hebei North University, Zhangjiakou, Hebei, China.
J Bacteriol. 2025 Jan 31;207(1):e0027524. doi: 10.1128/jb.00275-24. Epub 2024 Dec 26.
is an obligate intracellular bacterium of eukaryotic cells characterized by a unique biphasic life cycle; its biosynthesis and replication must occur within a cytoplasmic vacuole or inclusion. Certain inclusion membrane proteins have been demonstrated to mediate the interactions between intra-inclusion chlamydial organisms and the host cell. It has been demonstrated previously that the -encoded Cpn0308 localizes to the inclusion membrane; however, its function remains unknown. In the current study, a yeast two-hybrid assay was conducted to screen Cpn0308 as a bait against a HeLa cell cDNA library, revealing its binding to the host protein acyl-coenzyme A binding domain-containing 3 (ACBD3). The interaction between Cpn0308 and ACBD3 was confirmed through co-immunoprecipitation and GST (Glutathione S-transferase) pull-down assays. The two proteins were also co-localized in HeLa cells co-expressing Cpn0308 and ACBD3, as well as in -infected cells, as observed under confocal fluorescence microscopy. Given that ACBD3 plays a crucial role in maintaining host cell lipid homeostasis and its Golgi dynamic domain is responsible for interacting with Cpn0308, we hypothesize that the Cpn0308-ACBD3 interaction may facilitate 's acquisition of host lipids, thereby benefiting chlamydial survival. This study lays a foundation for further elucidating the mechanisms of Cpn0308-mediated pathogenesis.IMPORTANCEThe biosynthesis and replication of () must occur within the cytoplasmic vacuoles or inclusions of host cells. Inclusion bodies play a crucial role in mediating the interactions between and host cells. Cpn0308 is localized to the inclusion membrane; however, its function is unknown. In this study, Cpn0308 was found to bind to host protein acyl-coenzyme A binding domain-containing 3 (ACBD3) through some standard approaches. Co-localization of the two proteins was observed in both original HeLa cells and Cpn-infected HeLa cells. ACBD3 plays a significant role in maintaining lipid homeostasis in host cells; we speculate that the Cpn0308-ACBD3 interaction may facilitate the acquisition of host lipids by , thereby enhancing chlamydial survival.
是一种真核细胞内的专性寄生菌，具有独特的双相生命周期；其生物合成和复制必须在细胞质空泡或包涵体内进行。某些包涵体膜蛋白已被证明可介导包涵体内衣原体生物与宿主细胞之间的相互作用。先前已证明编码的Cpn0308定位于包涵体膜；然而，其功能仍然未知。在当前研究中，进行了酵母双杂交试验，以Cpn0308作为诱饵筛选HeLa细胞cDNA文库，发现它与宿主蛋白酰基辅酶A结合结构域包含蛋白3（ACBD3）结合。通过共免疫沉淀和GST（谷胱甘肽S-转移酶）下拉试验证实了Cpn0308与ACBD3之间的相互作用。在共表达Cpn0308和ACBD3的HeLa细胞以及感染衣原体的细胞中，通过共聚焦荧光显微镜观察到这两种蛋白也共定位。鉴于ACBD3在维持宿主细胞脂质稳态中起关键作用，并且其高尔基体动态结构域负责与Cpn0308相互作用，我们推测Cpn0308-ACBD3相互作用可能促进衣原体获取宿主脂质，从而有利于衣原体存活。本研究为进一步阐明Cpn0308介导的发病机制奠定了基础。重要性衣原体的生物合成和复制必须在宿主细胞的细胞质空泡或包涵体内进行。包涵体在介导衣原体与宿主细胞之间的相互作用中起关键作用。Cpn0308定位于包涵体膜；然而，其功能未知。在本研究中，通过一些标准方法发现Cpn0308与宿主蛋白酰基辅酶A结合结构域包含蛋白3（ACBD3）结合。在原始HeLa细胞和感染衣原体的HeLa细胞中均观察到这两种蛋白的共定位。ACBD3在维持宿主细胞脂质稳态中起重要作用；我们推测Cpn0308-ACBD3相互作用可能促进衣原体获取宿主脂质，从而增强衣原体存活能力。
