PMID- 31078419
OWN - NLM
STAT- MEDLINE
DCOM- 20200129
LR  - 20200129
IS  - 1873-281X (Electronic)
IS  - 1472-9792 (Linking)
VI  - 116S
DP  - 2019 May
TI  - Utility of OhrR-Ohr system for the expression of recombinant proteins in 
      mycobacteria and for the delivery of M. tuberculosis antigens to the phagosomal 
      compartment.
PG  - S19-S27
LID - S1472-9792(19)30146-5 [pii]
LID - 10.1016/j.tube.2019.04.007 [doi]
AB  - We have recently reported that in vitro and intracellular organic peroxide stress 
      oxidizes OhrR of Mycobacterium smegmatis and that the oxidized OhrR consequently 
      derepresses the expression of Ohr. Here we demonstrate that the OhrR-Ohr system 
      is highly useful for the expression of recombinant mycobacterial proteins and 
      also for the delivery of Mycobacterium tuberculosis (Mtb) antigens to the 
      phagosomal compartments. Recombinant M. smegmatis strains, which bear plasmid 
      constructs to express Ohr(2)-T85BCFP and Ohr(2)-MtrA, showed expression of fusion 
      proteins upon induction with t-butyl hydroperoxide (t-BHP) in a dose dependent 
      manner. The M. smegmatis expressed Ohr(2)-T85BCFP fusion could be affinity 
      purified by adding a 9x histidine tag to the C-terminal end of the fusion 
      protein. Further, mouse bone marrow derived macrophages (BMDMs) infected with 
      either recombinant M. smegmatis or BCG strains with ohr(2)-T85BCFP construct 
      showed expression of T85BCFP protein without any exogenously added inducer. In 
      addition, BMDMs infected with either recombinant BCG or Mtb with ohr(2)-T85BCFP 
      construct could effectively deliver the antigens to T-cells at higher levels than 
      strains bearing the control plasmid alone. Altogether, these results suggest that 
      the OhrR-Ohr system is a novel inducible system to study the biology and 
      pathogenesis of mycobacteria.
CI  - Copyright (c) 2019 Elsevier Ltd. All rights reserved.
FAU - Das, Kishore
AU  - Das K
AD  - Center of Emphasis in Infectious Diseases and Department of Biomedical Sciences, 
      Paul L. Foster School of Medicine, Texas Tech University Health Sciences Center 
      El Paso, El Paso, TX, 79905, USA.
FAU - Garnica, Omar
AU  - Garnica O
AD  - Center of Emphasis in Infectious Diseases and Department of Biomedical Sciences, 
      Paul L. Foster School of Medicine, Texas Tech University Health Sciences Center 
      El Paso, El Paso, TX, 79905, USA.
FAU - Dhandayuthapani, Subramanian
AU  - Dhandayuthapani S
AD  - Center of Emphasis in Infectious Diseases and Department of Biomedical Sciences, 
      Paul L. Foster School of Medicine, Texas Tech University Health Sciences Center 
      El Paso, El Paso, TX, 79905, USA; Graduate School of Biomedical Sciences, Texas 
      Tech University Health Sciences Center El Paso, El Paso, TX 79905, USA. 
      Electronic address: s.dhandayuthapani@ttuhsc.edu.
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
DEP - 20190427
PL  - Scotland
TA  - Tuberculosis (Edinb)
JT  - Tuberculosis (Edinburgh, Scotland)
JID - 100971555
RN  - 0 (ATP-Binding Cassette Transporters)
RN  - 0 (Antigens, Bacterial)
RN  - 0 (Bacterial Proteins)
RN  - 0 (MtrA protein, Bacteria)
RN  - 0 (Oligopeptides)
RN  - 0 (Recombinant Fusion Proteins)
RN  - 0 (Repressor Proteins)
RN  - 0 (peroxide repressor proteins)
RN  - 144058-44-6 (Mycobacterium tuberculosis antigens)
RN  - 4QD397987E (Histidine)
SB  - IM
MH  - ATP-Binding Cassette Transporters/genetics/metabolism
MH  - Animals
MH  - Antigens, Bacterial/genetics/*metabolism
MH  - Bacterial Proteins/genetics/*metabolism
MH  - Cells, Cultured
MH  - Gene Expression Regulation, Bacterial
MH  - Histidine/metabolism
MH  - Macrophages/metabolism/microbiology
MH  - Mice, Inbred C57BL
MH  - Mycobacterium bovis/genetics/metabolism
MH  - Mycobacterium smegmatis/genetics/*metabolism
MH  - Mycobacterium tuberculosis/genetics/*metabolism
MH  - Oligopeptides/metabolism
MH  - Phagosomes/genetics/*metabolism
MH  - Protein Transport
MH  - Recombinant Fusion Proteins/metabolism
MH  - Repressor Proteins/genetics/*metabolism
OTO - NOTNLM
OT  - Antigen
OT  - Delivery
OT  - Expression
OT  - Inducible
OT  - Macrophages
OT  - Peroxide
OT  - Phagosomes
OT  - Protein
OT  - Regulator
EDAT- 2019/05/13 06:00
MHDA- 2020/01/30 06:00
CRDT- 2019/05/13 06:00
PHST- 2018/11/19 00:00 [received]
PHST- 2018/11/19 00:00 [accepted]
PHST- 2019/05/13 06:00 [pubmed]
PHST- 2020/01/30 06:00 [medline]
PHST- 2019/05/13 06:00 [entrez]
AID - S1472-9792(19)30146-5 [pii]
AID - 10.1016/j.tube.2019.04.007 [doi]
PST - ppublish
SO  - Tuberculosis (Edinb). 2019 May;116S:S19-S27. doi: 10.1016/j.tube.2019.04.007. 
      Epub 2019 Apr 27.