title: Generation and envelope protein analysis of internalization defective Brucella abortus mutants in professional phagocytes, RAW 264.7.
abstract: Brucella abortus is a facultative intracellular bacteria that replicates within a macrophage without producing any classical virulence factors. It can become internalized to cells by zipper-like and/or swimming internalization mechanisms. However, the bacterial proteins involved in internalization remain unclear. To define these bacterial proteins, random insertion mutants of B. abortus were generated by the Tn5 transposome complexes. In all, 132 mutants were screened, cellular internalization-defective mutants were selected, and these genomic and envelope proteomic features were identified. The transposon insertion sites were ccmC,ppk and BruAb2_0168 for the mutant C10, C29 and D7, respectively. Mutant C10 showed a deficiency in internalization without any changes in expression of the cell envelope proteins; however, mutant C29 showed a reduced expression of OMP25, and a mutant D7 also showed reduced expression of OMP25, OMP28 and Porin2b. These results suggest OMP25 is not an essential factor, but might be involved in host cellular internalization. We identified the ppk gene and BruAb2_0168 locus which are associated to expression of OMP25, OMP28 and Porin2b as well as pleiotropic effects of ccmC gene.
authors: Department of Infectious Diseases, College of Veterinary Medicine, KRF Zoonotic Disease Priority Research Institute, Brain Korea 21 for Veterinary Science, Seoul National University, Seoul, South Korea.
authors: Seung Bin
authors: SB
authors: Cha
authors: Nabin
authors: N
authors: Rayamajhi
authors: Won Jung
authors: WJ
authors: Lee
authors: Min Kyung
authors: MK
authors: Shin
authors: Myung Hwan
authors: MH
authors: Jung
authors: Seung Won
authors: SW
authors: Shin
authors: Jong Wan
authors: JW
authors: Kim
authors: Han Sang
authors: HS
authors: Yoo
mesh_terms: D000818:Animals; D015139:Blotting, Southern; D002003:Brucella abortus / Q000235:genetics / Q000378:metabolism / Q000502:physiology*; D002460:Cell Line; D016521:Electrophoresis, Gel, Pulsed-Field; D015180:Electrophoresis, Gel, Two-Dimensional; D005798:Genes, Bacterial; D054884:Host-Pathogen Interactions; D008264:Macrophages / Q000382:microbiology*; D008565:Membrane Proteins / Q000235:genetics*; D051379:Mice; D009154:Mutation; D010586:Phagocytes / Q000382:microbiology*; D017856:Phosphotransferases (Phosphate Group Acceptor) / Q000235:genetics; D018272:Porins / Q000378:metabolism
chemical_list: D008565:Membrane Proteins; D018272:Porins; D017856:Phosphotransferases (Phosphate Group Acceptor); C022424:polyphosphate kinase