Protective Functions of Group 3 Late Embryogenesis Abundant (G3LEA) Proteins in Enterococcus faecium During Vancomycin T
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ORIGINAL RESEARCH ARTICLE
Protective Functions of Group 3 Late Embryogenesis Abundant (G3LEA) Proteins in Enterococcus faecium During Vancomycin Treatment Ahran Song1 • Boyong Kim1,2,3 • Eunyoung Kim1 • Jae Hwi Sung1 • Yoonjin Park1 • Sohyeon Park1 • Taegun Park1 • Jin Kwan Kim4 • Yoonhwa Jeong5,6 • Seung Gwan Lee1
Received: 11 March 2020 / Accepted: 10 July 2020 Ó Association of Microbiologists of India 2020
Abstract Late embryogenesis abundant (LEA) proteins protect organisms from various environmental stresses; however, the underlying mechanism of LEA mediated therapeutic evasion is still unclear in both eukaryotes and prokaryotes. In this study, group 3 LEA protein (G3LEA) of vancomycin-resistant Enterococcus faecium under sublethal concentration of vancomycin stress was evaluated and shown to have two functions: the first is the reduction of reactive oxygen species (ROS) content, preventing apoptosis by suppressing apoptotic proteins Cas3 and MAOB, and the second is activating specific drug efflux pumps. Sublethal vancomycin model was established with using Propidium Iodide (PI) stain. Real-time PCR was conducted to evaluate the expression of G3lea. Flow
cytometry and confocal microscope using Anti- G3LEA, anti- MAOB, and anti- Cas3 were performed to assess the expression of G3LEA. Under sublethal vancomycin stress, G3LEA is upregulated, suppressing the expression of apoptotic markers and increasing specific efflux markers. These results suggest that G3LEA protein suppresses antibiotic mediated apoptosis in prokaryotic cells and plays a key role in understanding and preventing antibiotic resistance. Keywords LEA proteins Vancomycin Enterococcus faecium Apoptosis ROS Efflux pump
Introduction Ahran Song and Boyong Kim have contributed equally to this work. & Seung Gwan Lee [email protected] 1
Department of Integrated Biomedical and Life Science, College of Health Science, Korea University, 145 Anam-ro, Seongbuk-gu, Seoul, South Korea
2
Life Together, 13 Gongdan-ro, Chuncheon-siGangwon, South Korea
3
Mitosbio, 13, Gongdan-ro, Chuncheon-si, Gangwon, South Korea
4
Department of Biomedical Laboratory Science, College of Health Science, Jungwon University, 85 Munmu-ro, Goesaneup, Goesan-gun, Chungbuk, South Korea
5
Research Center for Industrialization of Natural Nutraceuticals, Dankook University, Cheonan, Chungnam, Korea
6
Department of Food Science and Nutrition, College of Natural Science, Dankook University, Cheonan, Chungnam, Korea
Late embryogenesis abundant (LEA) proteins protect organisms from various abiotic and biotic stresses [1]. LEA proteins help plants endure desiccation, salinity changes, cold temperatures, and oxidative stress. LEA proteins were first discovered in cotton seeds (Gossypium hirsutum) [2]. Previous studies have revealed that LEA proteins are present not only in plants like Arabidopsis thaliana [3] but also in animals [4, 5] and microbes [6]. However, the exact mechanism by which LEA proteins effect this protection remains unclear. Enterococci are gram-posi
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