title: OtpR regulated the growth, cell morphology of B. melitensis and tolerance to β-lactam agents.
abstract: The intracellular pathogen, Brucella melitensis, possesses an operon with two components: otpR (BMEI0066), which encodes a response regulator, and BMEI0067, which encodes a putative cAMP-dependent protein kinase regulatory subunit. Previous studies have shown that a polar mutation in the BMEI0066 gene significantly decreased virulence and stress tolerance in Brucella. In this study, we constructed non-polar mutant with deletion of otpR, as well as its complementary strain to further investigate the function of otpR. The ΔotpR mutant produced smaller colonies on TSA plates, and grew slower in tryptic soy broth compared to 16M or the otpR-complemented strain CotpR. Electron microscopy revealed that ΔotpR displayed an unusual, irregular deformation of the cell surface in contrast to the native coccobacillus shape of 16M. These results showed that OtpR played a key role in the maintenance of cell shape. To determine the effect of the otpR mutant on antibiotic susceptibility, compared the parent strain, the mutant was two- to eight-fold more susceptible to all the β-lactam antibiotics tested. Furthermore, comparative real-time qPCR of genes that related to penicillin binding proteins of cell wall synthesis and cell division showed that the otpR mutation resulted in reduced expression of pbp1C, pbp6B, pbp6C and ftsQ. Taken together, these data revealed that the OtpR activity is necessary for growth, and cell morphology and tolerance to β-lactam agents of B. melitensis.
authors.affiliation: Key Laboratory of Animal Epidemiology and Zoonosis of Ministry of Agriculture, College of Veterinary Medicine, China Agricultural University, Beijing 100193, People's Republic of China.
authors.forename: Wenjuan
authors.initials: W
authors.lastname: Liu
authors.forename: Hao
authors.initials: H
authors.lastname: Dong
authors.forename: Wenxiao
authors.initials: W
authors.lastname: Liu
authors.forename: Xiaolei
authors.initials: X
authors.lastname: Gao
authors.forename: Chunyan
authors.initials: C
authors.lastname: Zhang
authors.forename: Qingmin
authors.initials: Q
authors.lastname: Wu
mesh_terms: D000900:Anti-Bacterial Agents / Q000494:pharmacology*; D001426:Bacterial Proteins / Q000235:genetics* / Q000378:metabolism*; D017347:Brucella melitensis / Q000187:drug effects* / Q000235:genetics* / Q000378:metabolism / Q000648:ultrastructure; D004361:Drug Tolerance / Q000235:genetics; D011112:Polymyxin B / Q000494:pharmacology; D017384:Sequence Deletion; D047090:beta-Lactams / Q000494:pharmacology*
chemical_list: D000900:Anti-Bacterial Agents; D001426:Bacterial Proteins; D047090:beta-Lactams; D011112:Polymyxin B