Effects of the Ccdc115 gene on the phagosome acidification and viability of RAW264.7 cells infected with Salmonella Typhimurium
10.3969/j.issn.1002-2694.2025.00.101
- VernacularTitle:Ccdc115基因对鼠伤寒沙门菌感染RAW264.7细胞吞噬体酸化及生存能力的影响
- Author:
Rong-xian XIE
1
;
Long-yun CHENG
1
;
Xi-lu YUAN
1
;
Li LI
1
;
Bing-qing LI
1
;
Hai-hong JIA
1
Author Information
1. 山东第一医科大学(山东省医学科学院)临床与基础医学院(基础医学研究所),济南 250062
- Publication Type:Journal Article
- Keywords:
Coiled-Coil Domain-Containing Protein 115;
Salmonella Typhimurium;
RAW264.7;
intracellular survival;
phago-some acidification
- From:
Chinese Journal of Zoonoses
2025;41(6):559-566
- CountryChina
- Language:Chinese
-
Abstract:
This work was aimed at analyzing the protein characteristics of Coiled-Coil Domain-Containing Protein 115(CCDC115)and using Ccdc115-deficient mouse monocyte-macrophage leukemia cells(RAW264.7)to explore the influence of CCDC115 on the intracellular survival of Salmonella Typhimurium.Bioinformatics analysis was conducted to examine the fundamental attributes of CCDC115,which was determined to be an unstable protein consisting of two α-helices and an intervening disordered re-gion,devoid of any transmembrane structural domains.A RAW264.7-Ccdc115-KO cell line was successfully established with CRISPR/Cas9 gene-editing technology.To elucidate the effects of CCDC115 on the intracellular survival of Salmonella Typhimurium,we infected RAW264.7 cells with Salmonella Typhimurium.The expression of CCDC115 was found to be upregulated at both the mRNA and protein levels post-infection,according to RT-qPCR and western blot analysis.Via counting of colony-forming units(CFU),the proliferation rate of Salmonella Typhimurium within RAW264.7-Ccdc115-KO cells was found to be 1.5-fold higher than that in RAW264.7 cells.Acidification imaging studies indicated that,whereas Salmonella Typhimurium phagosomes underwent acidifi-cation in RAW264.7 cells,this process was absent in RAW264.7-Ccdc115-KO cells.In conclusion,the study successfully estab-lished a RAW264.7-Ccdc115-KO cell line and demonstrated that the expression of CCDC115 is elevated during Salmonella Ty-phimurium infection,thus potentially inhibiting the intracellular survival of Salmonella Typhimurium by facilitating phagosome acidifi-cation.This study lay a theoretical foundation for functional studies of CCDC115 and the investigation of mechanisms regulating the survival of intracellular Salmonella Typhimurium.
