Galectin-3 inhibits galectin-8/parkin- mediated ubiquitination of group A streptococcus

Yi Lin Cheng, Yan Wei Wu, Chih Feng Kuo, Shiou Ling Lu, Fu-Tong Liu, Robert Anderson, Chiou Feng Lin, Yi Ling Liu, Wan Yu Wang, Ying Da Chen, Po Xing Zheng, Jiunn Jong Wu, Yee Shin Lin

Research output: Contribution to journalArticle

4 Citations (Scopus)

Abstract

Group A streptococcus (GAS) is an important human pathogen that causes a wide variety of cutaneous and systemic infections. Although originally thought to be an extracellular bacterium, numerous studies have demonstrated that GAS can trigger internalization into nonimmune cells to escape from immune surveillance or antibiotic-mediated killing. Epithelial cells possess a defense mechanism involving autophagy-mediated targeting and killing of GAS within lysosome-fused autophagosomes. In endothelial cells, in contrast, we previously showed that autophagy is not sufficient for GAS killing. In the present study, we showed higher galectin-3 (Gal-3) expression and lower Gal-8 expression in endothelial cells than in epithelial cells. The recruitment of Gal-3 to GAS is higher and the recruitment of Gal-8 to GAS is lower in endothelial cells than in epithelial cells. We further showed that Gal-3 promotes GAS replication and diminishes the recruitment of Gal-8 and ubiquitin, the latter of which is a critical protein for autophagy sequestration. After knockdown of Gal-3 in endothelial cells, the colocalization of Gal-8, parkin, and ubiquitin-decorated GAS is significantly increased, as is the interaction of Gal-8 and parkin, an E3 ligase. Furthermore, inhibition of Gal-8 in epithelial cells attenuates recruitment of parkin; both Gal-8 and parkin contribute to ubiquitin recruitment and GAS elimination. Animal studies confirmed that Gal-3-knockout mice develop less-severe skin damage and that GAS replication can be detected only in the air pouch and not in organs and endothelial cells. These results demonstrate that Gal-3 inhibits ubiquitin recruitment by blocking Gal-8 and parkin recruitment, resulting in GAS replication in endothelial cells. IMPORTANCE In epithelial cells, GAS can be efficiently killed within the lysosomefused autophaosome compartment. However, we previously showed that, in spite of LC-3 recruitment, the autophagic machinery is not sufficient for GAS killing in endothelial cells. In this report, we provide the first evidence that Gal-3, highly expressed in endothelial cells, blocks the tagging of ubiquitin to GAS by inhibiting recruitment of Gal-8 and parkin, leading to an enhancement of GAS replication. We also provide the first demonstration that Gal-8 can interact with parkin, the critical E3 ligase, for resistance to intracellular bacteria by facilitating the decoration of bacteria with ubiquitin chains. Our findings reveal that differential levels of Gal-3 and Gal-8 expression and recruitment to GAS between epithelial cells and endothelial cells may contribute to the different outcomes of GAS elimination or survival and growth of GAS in these two types of cells.

Original languageEnglish (US)
Article numbere00899-17
JournalmBio
Volume8
Issue number4
DOIs
StatePublished - Jul 1 2017
Externally publishedYes

Fingerprint

Galectins
Galectin 3
Ubiquitination
Streptococcus
Endothelial Cells
Ubiquitin
Epithelial Cells
Autophagy
Ubiquitin-Protein Ligases
Bacteria
Skin

Keywords

  • Galectin-3
  • Galectin-8
  • Group A streptococcus
  • Parkin
  • Ubiquitin

ASJC Scopus subject areas

  • Microbiology
  • Virology

Cite this

Cheng, Y. L., Wu, Y. W., Kuo, C. F., Lu, S. L., Liu, F-T., Anderson, R., ... Lin, Y. S. (2017). Galectin-3 inhibits galectin-8/parkin- mediated ubiquitination of group A streptococcus. mBio, 8(4), [e00899-17]. https://doi.org/10.1128/mBio.00899-17

Galectin-3 inhibits galectin-8/parkin- mediated ubiquitination of group A streptococcus. / Cheng, Yi Lin; Wu, Yan Wei; Kuo, Chih Feng; Lu, Shiou Ling; Liu, Fu-Tong; Anderson, Robert; Lin, Chiou Feng; Liu, Yi Ling; Wang, Wan Yu; Chen, Ying Da; Zheng, Po Xing; Wu, Jiunn Jong; Lin, Yee Shin.

In: mBio, Vol. 8, No. 4, e00899-17, 01.07.2017.

Research output: Contribution to journalArticle

Cheng, YL, Wu, YW, Kuo, CF, Lu, SL, Liu, F-T, Anderson, R, Lin, CF, Liu, YL, Wang, WY, Chen, YD, Zheng, PX, Wu, JJ & Lin, YS 2017, 'Galectin-3 inhibits galectin-8/parkin- mediated ubiquitination of group A streptococcus', mBio, vol. 8, no. 4, e00899-17. https://doi.org/10.1128/mBio.00899-17
Cheng YL, Wu YW, Kuo CF, Lu SL, Liu F-T, Anderson R et al. Galectin-3 inhibits galectin-8/parkin- mediated ubiquitination of group A streptococcus. mBio. 2017 Jul 1;8(4). e00899-17. https://doi.org/10.1128/mBio.00899-17
Cheng, Yi Lin ; Wu, Yan Wei ; Kuo, Chih Feng ; Lu, Shiou Ling ; Liu, Fu-Tong ; Anderson, Robert ; Lin, Chiou Feng ; Liu, Yi Ling ; Wang, Wan Yu ; Chen, Ying Da ; Zheng, Po Xing ; Wu, Jiunn Jong ; Lin, Yee Shin. / Galectin-3 inhibits galectin-8/parkin- mediated ubiquitination of group A streptococcus. In: mBio. 2017 ; Vol. 8, No. 4.
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abstract = "Group A streptococcus (GAS) is an important human pathogen that causes a wide variety of cutaneous and systemic infections. Although originally thought to be an extracellular bacterium, numerous studies have demonstrated that GAS can trigger internalization into nonimmune cells to escape from immune surveillance or antibiotic-mediated killing. Epithelial cells possess a defense mechanism involving autophagy-mediated targeting and killing of GAS within lysosome-fused autophagosomes. In endothelial cells, in contrast, we previously showed that autophagy is not sufficient for GAS killing. In the present study, we showed higher galectin-3 (Gal-3) expression and lower Gal-8 expression in endothelial cells than in epithelial cells. The recruitment of Gal-3 to GAS is higher and the recruitment of Gal-8 to GAS is lower in endothelial cells than in epithelial cells. We further showed that Gal-3 promotes GAS replication and diminishes the recruitment of Gal-8 and ubiquitin, the latter of which is a critical protein for autophagy sequestration. After knockdown of Gal-3 in endothelial cells, the colocalization of Gal-8, parkin, and ubiquitin-decorated GAS is significantly increased, as is the interaction of Gal-8 and parkin, an E3 ligase. Furthermore, inhibition of Gal-8 in epithelial cells attenuates recruitment of parkin; both Gal-8 and parkin contribute to ubiquitin recruitment and GAS elimination. Animal studies confirmed that Gal-3-knockout mice develop less-severe skin damage and that GAS replication can be detected only in the air pouch and not in organs and endothelial cells. These results demonstrate that Gal-3 inhibits ubiquitin recruitment by blocking Gal-8 and parkin recruitment, resulting in GAS replication in endothelial cells. IMPORTANCE In epithelial cells, GAS can be efficiently killed within the lysosomefused autophaosome compartment. However, we previously showed that, in spite of LC-3 recruitment, the autophagic machinery is not sufficient for GAS killing in endothelial cells. In this report, we provide the first evidence that Gal-3, highly expressed in endothelial cells, blocks the tagging of ubiquitin to GAS by inhibiting recruitment of Gal-8 and parkin, leading to an enhancement of GAS replication. We also provide the first demonstration that Gal-8 can interact with parkin, the critical E3 ligase, for resistance to intracellular bacteria by facilitating the decoration of bacteria with ubiquitin chains. Our findings reveal that differential levels of Gal-3 and Gal-8 expression and recruitment to GAS between epithelial cells and endothelial cells may contribute to the different outcomes of GAS elimination or survival and growth of GAS in these two types of cells.",
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AU - Cheng, Yi Lin

AU - Wu, Yan Wei

AU - Kuo, Chih Feng

AU - Lu, Shiou Ling

AU - Liu, Fu-Tong

AU - Anderson, Robert

AU - Lin, Chiou Feng

AU - Liu, Yi Ling

AU - Wang, Wan Yu

AU - Chen, Ying Da

AU - Zheng, Po Xing

AU - Wu, Jiunn Jong

AU - Lin, Yee Shin

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N2 - Group A streptococcus (GAS) is an important human pathogen that causes a wide variety of cutaneous and systemic infections. Although originally thought to be an extracellular bacterium, numerous studies have demonstrated that GAS can trigger internalization into nonimmune cells to escape from immune surveillance or antibiotic-mediated killing. Epithelial cells possess a defense mechanism involving autophagy-mediated targeting and killing of GAS within lysosome-fused autophagosomes. In endothelial cells, in contrast, we previously showed that autophagy is not sufficient for GAS killing. In the present study, we showed higher galectin-3 (Gal-3) expression and lower Gal-8 expression in endothelial cells than in epithelial cells. The recruitment of Gal-3 to GAS is higher and the recruitment of Gal-8 to GAS is lower in endothelial cells than in epithelial cells. We further showed that Gal-3 promotes GAS replication and diminishes the recruitment of Gal-8 and ubiquitin, the latter of which is a critical protein for autophagy sequestration. After knockdown of Gal-3 in endothelial cells, the colocalization of Gal-8, parkin, and ubiquitin-decorated GAS is significantly increased, as is the interaction of Gal-8 and parkin, an E3 ligase. Furthermore, inhibition of Gal-8 in epithelial cells attenuates recruitment of parkin; both Gal-8 and parkin contribute to ubiquitin recruitment and GAS elimination. Animal studies confirmed that Gal-3-knockout mice develop less-severe skin damage and that GAS replication can be detected only in the air pouch and not in organs and endothelial cells. These results demonstrate that Gal-3 inhibits ubiquitin recruitment by blocking Gal-8 and parkin recruitment, resulting in GAS replication in endothelial cells. IMPORTANCE In epithelial cells, GAS can be efficiently killed within the lysosomefused autophaosome compartment. However, we previously showed that, in spite of LC-3 recruitment, the autophagic machinery is not sufficient for GAS killing in endothelial cells. In this report, we provide the first evidence that Gal-3, highly expressed in endothelial cells, blocks the tagging of ubiquitin to GAS by inhibiting recruitment of Gal-8 and parkin, leading to an enhancement of GAS replication. We also provide the first demonstration that Gal-8 can interact with parkin, the critical E3 ligase, for resistance to intracellular bacteria by facilitating the decoration of bacteria with ubiquitin chains. Our findings reveal that differential levels of Gal-3 and Gal-8 expression and recruitment to GAS between epithelial cells and endothelial cells may contribute to the different outcomes of GAS elimination or survival and growth of GAS in these two types of cells.

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