A comprehensive proteogenomic study of the human Brucella vaccine strain 104 M

BMC Genomics

Research Article

Kun Liu¹  Mengying Qian¹  Dayong Dong¹  Xiaodong Zai¹  Ying Yin¹  Yaohui Li¹  Junjie Xu¹  Shanhu Li¹  Wei Chen¹  Taoran Zhao¹  Ling Fu¹  Qiaoling Yang¹  Ruihua Li¹ 

[1] Laboratory of Vaccine and Antibody Engineering, Beijing Institute of Biotechnology, Beijing, China;

关键词: Brucella abortus;    Proteogenomic;    Genome reannotation;    Virulence factor;    Protective antigen;    Multichromosomal bacteria;   

DOI  :  10.1186/s12864-017-3800-9

 received in 2016-11-21, accepted in 2017-05-16,  发布年份 2017

来源: Springer

【 摘 要 】

BackgroundBrucella spp. are Gram-negative, facultative intracellular pathogens that cause brucellosis in both humans and animals. The B. abortus vaccine strain 104 M is the only vaccine available in China for the prevention of brucellosis in humans. Although the B. abortus 104 M genome has been fully sequenced, the current genome annotations are not yet complete. In addition, the main mechanisms underpinning its residual toxicity and vaccine-induced immune protection have yet to be elucidated. Mapping the proteome of B. abortus 104 M will help to improve genome annotation quality, thereby facilitating a greater understanding of its biology.ResultsIn this study, we utilized a proteogenomic approach that combined subcellular fractionation and peptide fractionation to perform a whole-proteome analysis and genome reannotation of B. abortus 104 M using high-resolution mass spectrometry. In total, 1,729 proteins (56.3% of 3,072) including 218 hypothetical proteins were identified using the culture conditions that were employed this study. The annotations of the B. abortus 104 M genome were also refined following identification and validation by reverse transcription-PCR. In addition, 14 pivotal virulence factors and 17 known protective antigens known to be involved in residual toxicity and immune protection were confirmed at the protein level following induction by the 104 M vaccine. Moreover, a further insight into the cell biology of multichromosomal bacteria was obtained following the elucidation of differences in protein expression levels between the small and large chromosomes.ConclusionsThe work presented in this report used a proteogenomic approach to perform whole-proteome analysis and genome reannotation in B. abortus 104 M; this work helped to improve genome annotation quality. Our analysis of virulence factors, protective antigens and other protein effectors provided the basis for further research to elucidate the mechanisms of residual toxicity and immune protection induced by the 104 M vaccine. Finally, the potential link between replication dynamics, gene function, and protein expression levels in this multichromosomal bacterium was detailed.

【 授权许可】

CC BY   
© The Author(s). 2017