Lethality of MalE-LacZ hybrid protein shares mechanistic attributes with oxidative component of antibiotic lethality
Downstream metabolic events can contribute to the lethality of drugs or agents that interact with a primary cellular target. In bacteria, the production of reactive oxygen species (ROS) has been associated with the lethal effects of a variety of stresses including bactericidal antibiotics, but the r...
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| Format: | Article |
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National Academy of Sciences (U.S.)
2018
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| Online Access: | http://hdl.handle.net/1721.1/114885 https://orcid.org/0000-0002-7127-9833 https://orcid.org/0000-0003-0921-4657 https://orcid.org/0000-0003-4710-1389 https://orcid.org/0000-0002-5560-8246 https://orcid.org/0000-0001-7243-8261 |
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| author | Liu, Xiaobo Andreescu, Silvana Takahashi, Noriko Gruber, Charley C Yang, Jason Hung-Ying Braff, Dana Yashaswini, Chittampalli N. Bhubhanil, Sakkarin Furuta, Yoshikazu Collins, James J. Walker, Graham C. |
| author2 | Massachusetts Institute of Technology. Institute for Medical Engineering & Science |
| author_facet | Massachusetts Institute of Technology. Institute for Medical Engineering & Science Liu, Xiaobo Andreescu, Silvana Takahashi, Noriko Gruber, Charley C Yang, Jason Hung-Ying Braff, Dana Yashaswini, Chittampalli N. Bhubhanil, Sakkarin Furuta, Yoshikazu Collins, James J. Walker, Graham C. |
| author_sort | Liu, Xiaobo |
| collection | MIT |
| description | Downstream metabolic events can contribute to the lethality of drugs or agents that interact with a primary cellular target. In bacteria, the production of reactive oxygen species (ROS) has been associated with the lethal effects of a variety of stresses including bactericidal antibiotics, but the relative contribution of this oxidative component to cell death depends on a variety of factors. Experimental evidence has suggested that unresolvable DNA problems caused by incorporation of oxidized nucleotides into nascent DNA followed by incomplete base excision repair contribute to the ROS-dependent component of antibiotic lethality. Expression of the chimeric periplasmic-cytoplasmic MalE-LacZ[subscript 72 – 47] protein is an historically important lethal stress originally identified during seminal genetic experiments that defined the SecY-dependent protein translocation system. Multiple, independent lines of evidence presented here indicate that the predominant mechanism for MalE-LacZ lethality shares attributes with the ROS-dependent component of antibiotic lethality. MalE-LacZ lethality requires molecular oxygen, and its expression induces ROS production. The increased susceptibility of mutants sensitive to oxidative stress to MalE-LacZ lethality indicates that ROS contribute causally to cell death rather than simply being produced by dying cells. Observations that support the proposed mechanism of cell death include MalE-LacZ expression being bacteriostatic rather than bactericidal in cells that over-express MutT, a nucleotide sanitizer that hydrolyzes 8-oxo-dGTP to the monophosphate, or that lack MutM and MutY, DNA glycosylases that process base pairs involving 8-oxo-dGTP. Our studies suggest stress-induced physiological changes that favor this mode of ROS-dependent death. |
| first_indexed | 2024-09-23T11:02:42Z |
| format | Article |
| id | mit-1721.1/114885 |
| institution | Massachusetts Institute of Technology |
| last_indexed | 2024-09-23T11:02:42Z |
| publishDate | 2018 |
| publisher | National Academy of Sciences (U.S.) |
| record_format | dspace |
| spelling | mit-1721.1/1148852024-03-20T19:42:34Z Lethality of MalE-LacZ hybrid protein shares mechanistic attributes with oxidative component of antibiotic lethality Liu, Xiaobo Andreescu, Silvana Takahashi, Noriko Gruber, Charley C Yang, Jason Hung-Ying Braff, Dana Yashaswini, Chittampalli N. Bhubhanil, Sakkarin Furuta, Yoshikazu Collins, James J. Walker, Graham C. Massachusetts Institute of Technology. Institute for Medical Engineering & Science Massachusetts Institute of Technology. Department of Biological Engineering Massachusetts Institute of Technology. Department of Biology Takahashi, Noriko Gruber, Charley C Yang, Jason Hung-Ying Braff, Dana Yashaswini, Chittampalli N. Bhubhanil, Sakkarin Furuta, Yoshikazu Collins, James J. Walker, Graham C Downstream metabolic events can contribute to the lethality of drugs or agents that interact with a primary cellular target. In bacteria, the production of reactive oxygen species (ROS) has been associated with the lethal effects of a variety of stresses including bactericidal antibiotics, but the relative contribution of this oxidative component to cell death depends on a variety of factors. Experimental evidence has suggested that unresolvable DNA problems caused by incorporation of oxidized nucleotides into nascent DNA followed by incomplete base excision repair contribute to the ROS-dependent component of antibiotic lethality. Expression of the chimeric periplasmic-cytoplasmic MalE-LacZ[subscript 72 – 47] protein is an historically important lethal stress originally identified during seminal genetic experiments that defined the SecY-dependent protein translocation system. Multiple, independent lines of evidence presented here indicate that the predominant mechanism for MalE-LacZ lethality shares attributes with the ROS-dependent component of antibiotic lethality. MalE-LacZ lethality requires molecular oxygen, and its expression induces ROS production. The increased susceptibility of mutants sensitive to oxidative stress to MalE-LacZ lethality indicates that ROS contribute causally to cell death rather than simply being produced by dying cells. Observations that support the proposed mechanism of cell death include MalE-LacZ expression being bacteriostatic rather than bactericidal in cells that over-express MutT, a nucleotide sanitizer that hydrolyzes 8-oxo-dGTP to the monophosphate, or that lack MutM and MutY, DNA glycosylases that process base pairs involving 8-oxo-dGTP. Our studies suggest stress-induced physiological changes that favor this mode of ROS-dependent death. National Institutes of Health (U.S.) (Grant R01CA021615) Defense Threat Reduction Agency (DTRA) (Grant HDTRA1-15-1-0051) National Science Foundation (U.S.) (Grant 1336493) National Institutes of Health (U.S.) (Grant K99GM118907) 2018-04-23T18:15:45Z 2018-04-23T18:15:45Z 2017-08 2017-05 2018-04-20T13:43:59Z Article http://purl.org/eprint/type/ConferencePaper 0027-8424 1091-6490 http://hdl.handle.net/1721.1/114885 Takahashi, Noriko et al. “Lethality of MalE-LacZ Hybrid Protein Shares Mechanistic Attributes with Oxidative Component of Antibiotic Lethality.” Proceedings of the National Academy of Sciences 114, 34 (August 2017): 9164–9169 © 2017 National Academy of Sciences https://orcid.org/0000-0002-7127-9833 https://orcid.org/0000-0003-0921-4657 https://orcid.org/0000-0003-4710-1389 https://orcid.org/0000-0002-5560-8246 https://orcid.org/0000-0001-7243-8261 http://dx.doi.org/10.1073/PNAS.1707466114 Proceedings of the National Academy of Sciences Article is made available in accordance with the publisher's policy and may be subject to US copyright law. Please refer to the publisher's site for terms of use. application/pdf National Academy of Sciences (U.S.) National Academy of Sciences |
| spellingShingle | Liu, Xiaobo Andreescu, Silvana Takahashi, Noriko Gruber, Charley C Yang, Jason Hung-Ying Braff, Dana Yashaswini, Chittampalli N. Bhubhanil, Sakkarin Furuta, Yoshikazu Collins, James J. Walker, Graham C. Lethality of MalE-LacZ hybrid protein shares mechanistic attributes with oxidative component of antibiotic lethality |
| title | Lethality of MalE-LacZ hybrid protein shares mechanistic attributes with oxidative component of antibiotic lethality |
| title_full | Lethality of MalE-LacZ hybrid protein shares mechanistic attributes with oxidative component of antibiotic lethality |
| title_fullStr | Lethality of MalE-LacZ hybrid protein shares mechanistic attributes with oxidative component of antibiotic lethality |
| title_full_unstemmed | Lethality of MalE-LacZ hybrid protein shares mechanistic attributes with oxidative component of antibiotic lethality |
| title_short | Lethality of MalE-LacZ hybrid protein shares mechanistic attributes with oxidative component of antibiotic lethality |
| title_sort | lethality of male lacz hybrid protein shares mechanistic attributes with oxidative component of antibiotic lethality |
| url | http://hdl.handle.net/1721.1/114885 https://orcid.org/0000-0002-7127-9833 https://orcid.org/0000-0003-0921-4657 https://orcid.org/0000-0003-4710-1389 https://orcid.org/0000-0002-5560-8246 https://orcid.org/0000-0001-7243-8261 |
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