Analysis of Brucella melitensis wbkC protein affecting the lncRNA expression profiles of RAW264.7 cells and mining of autophagy pathway lncRNAs

Background Brucellosis is a veterinary and human disease caused by Brucella , which not only causes serious public safety but also affects the development of animal husbandry and international trade. The formyl transferase encoded by the wbkC gene plays an important role in the synthesis of lipopolysaccharide (LPS), an important virulence factor of Brucella melitensis . Long-stranded non-coding RNAs (lncRNAs), as an emerging regulatory molecule, are thought to be extensively involved in the regulation of cellular activities. In this paper, by studying the analysis of Brucella melitensis . wbkC protein affecting the lncRNA expression profile of RAW264.7 cells and the mining of autophagy pathway lncRNAs in the sheep species, the aim of this study was to reveal the mechanism by which B. melitensis affects macrophage autophagy and intracellular survival. Methods In this study, B. melitensis wbkC protein was obtained by prokaryotic expression, and polyclonal antibody to wbkC protein was prepared by immunizing rabbits. Recombinant adenovirus for wbkC gene overexpression was prepared to mediate wbkC overexpression in RAW264.7 cells with Ad_wbkC. The wbkC protein was analyzed to affect the lncRNA expression profile of RAW264.7 cells by transcriptomics sequencing technology to mine the autophagy pathway-related lncRNAs. Results The results showed that Ad_wbkC successfully mediated the overexpression of wbkC in RAW264.7 cells, and a total of 32 expression up-regulated lncRNAs versus 30 expression down-regulated lncRNAs were identified in the Ad_wbkC group as compared with the control group. By analyzing the functional enrichment of the lncRNA target genes GO and KEGG, the lncRNAs with TOP15 expression related to autophagy were screened for qRT-PCR validation. The validation results showed that lncRNA 4933430A20Rik and lncRNA B930036N10Rik were consistent with the sequencing results. Conclusions B. melitensis wbkc protein further affects macrophage autophagy and intracellular survival by influencing lncRNA expression.