Abstract
Group A Streptococcus (GAS) is one of the major human pathogens that can invade nonphagocytic cells. GAS internalized through endocytosis secretes the pore-forming toxin Streptolysin O (SLO) to escape into the cytoplasm. The cytosolic GAS is selectively captured by autophagic membranes (GAS-containing autophagosome-like vacuoles, GcAVs) and delivered to lysosomes for degradation. Macroautophagy (referred to as autophagy hereafter) is a highly conserved lysosome-mediated catabolic process, which is critical for cellular homeostasis. Autophagy also acts as an intracellular immune system. In this section, we describe how to identify GcAVs in infected cells using fluorescent microscopy.
This is a preview of subscription content, log in via an institution to check access.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Walker MJ, Barnett TC, McArthur JD et al (2014) Disease manifestations and pathogenic mechanisms of group A Streptococcus. Clin Microbiol Rev 27:264–301
Greco R, De Martino L, Donnarumma G et al (1995) Invasion of cultured human cells by Streptococcus pyogenes. Res Microbiol 146:551–560
LaPenta D, Rubens C, Chi E, Cleary PP (1994) Group A streptococci efficiently invade human respiratory epithelial cells. Proc Natl Acad Sci U S A 91:12115–12119
Nakagawa I, Amano A, Mizushima N et al (2004) Autophagy defends cells against invading group A Streptococcus. Science 306:1037–1040
O'Seaghdha M, Wessels MR (2013) Streptolysin O and its co-toxin NAD-glycohydrolase protect group A Streptococcus from xenophagic killing. PLoS Pathog 9:e1003394
Barnett TC, Liebl D, Seymour LM et al (2013) The globally disseminated M1T1 clone of group A Streptococcus evades autophagy for intracellular replication. Cell Host Microbe 14:675–682
Mizushima N, Komatsu M (2011) Autophagy: renovation of cells and tissues. Cell 147:728–741
Gatica D, Lahiri V, Klionsky DJ (2018) Cargo recognition and degradation by selective autophagy. Nat Cell Biol 20:233–242
Mizushima N, Yoshimori T, Levine B (2010) Methods in mammalian autophagy research. Cell 140:313–326
Heckmann BL, Boada-Romero E, Cunha LD et al (2017) LC3-associated phagocytosis and inflammation. J Mol Biol 429:3561–3576
Sharma V, Verma S, Seranova E et al (2018) Selective autophagy and xenophagy in infection and disease. Front Cell Dev Biol 6:147
Lu SL, Kawabata T, Cheng YL et al (2017) Endothelial cells are intrinsically defective in xenophagy of Streptococcus pyogenes. PLoS Pathog 13:e1006444
Minowa-Nozawa A, Nozawa T, Okamoto-Furuta K et al (2017) Rab35 GTPase recruits NDP52 to autophagy targets. EMBO J 36:2790–2807
Joubert PE, Meiffren G, Grégoire IP et al (2009) Autophagy induction by the pathogen receptor CD46. Cell Host Microbe 6:354–366
Thurston TL, Ryzhakov G, Bloor S et al (2009) The TBK1 adaptor and autophagy receptor NDP52 restricts the proliferation of ubiquitin-coated bacteria. Nat Immunol 10:1215–1221
Toh H, Nozawa T, Minowa-Nozawa A et al (2019) Group A Streptococcus modulates RAB1- and PIK3C3 complex-dependent autophagy. Autophagy. https://doi.org/10.1080/15548627.2019.1628539
Nakajima S, Aikawa C, Nozawa T et al (2017) Bcl-xL affects group A Streptococcus-induced autophagy directly, by inhibiting fusion between autophagosomes and lysosomes, and indirectly, by inhibiting bacterial internalization via interaction with Beclin 1-UVRAG. PLoS One 12:e0170138
Nozawa T, Aikawa C, Goda A et al (2012) The small GTPases Rab9A and Rab23 function at distinct steps in autophagy during group A Streptococcus infection. Cell Microbiol 14:1149–1165
Haobam B, Nozawa T, Minowa-Nozawa A et al (2014) Rab17-mediated recycling endosomes contribute to autophagosome formation in response to group A Streptococcus invasion. Cell Microbiol 16:1806–1821
Nakajima K, Nozawa T, Minowa-Nozawa A et al (2019) RAB30 regulates PI4KB (phosphatidylinositol 4-kinase beta)-dependent autophagy against group A Streptococcus. Autophagy 15:466–477
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2020 Springer Science+Business Media, LLC, part of Springer Nature
About this protocol
Cite this protocol
Nozawa, T., Nakagawa, I. (2020). Identification of Group A Streptococcus-Containing Autophagosome-Like Vacuoles. In: Proft, T., Loh, J. (eds) Group A Streptococcus. Methods in Molecular Biology, vol 2136. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-0467-0_16
Download citation
DOI: https://doi.org/10.1007/978-1-0716-0467-0_16
Published:
Publisher Name: Humana, New York, NY
Print ISBN: 978-1-0716-0466-3
Online ISBN: 978-1-0716-0467-0
eBook Packages: Springer Protocols