GsdmD p30 elicited by caspase-11 during pyroptosis forms pores in membranes

Aglietti, Robin A.; Estevez, Alberto; Gupta, Aaron; Ramirez, Monica Gonzalez; Liu, Peter S.; Kayagaki, Nobuhiko; Ciferri, Claudio; Dixit, Vishva M.; Dueber, Erin C.

doi:10.1073/pnas.1607769113

Cited by 759 publications

(654 citation statements)

References 27 publications

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“…Importantly, our in vitro assays show that the formation of the GSDMD pore does not require other proteins as co‐factors or membrane receptors, demonstrating that GSDMD Nterm is the sole and final executor of pyroptotic cell death (Fig 7). While this manuscript was in revision, other reports have been published that confirm our findings that the N‐terminal fragment of GSDMD causes pyroptosis through the formation of a plasma membrane pore and that demonstrate that the N‐terminal domains of other gasdermin family members share this pore‐forming activity (Aglietti et al , 2016; Ding et al , 2016). …”

Section: Discussionsupporting

confidence: 79%

GSDMD membrane pore formation constitutes the mechanism of pyroptotic cell death

et al. 2016

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Pyroptosis is a lytic type of cell death that is initiated by inflammatory caspases. These caspases are activated within multi‐protein inflammasome complexes that assemble in response to pathogens and endogenous danger signals. Pyroptotic cell death has been proposed to proceed via the formation of a plasma membrane pore, but the underlying molecular mechanism has remained unclear. Recently, gasdermin D (GSDMD), a member of the ill‐characterized gasdermin protein family, was identified as a caspase substrate and an essential mediator of pyroptosis. GSDMD is thus a candidate for pyroptotic pore formation. Here, we characterize GSDMD function in live cells and in vitro. We show that the N‐terminal fragment of caspase‐1‐cleaved GSDMD rapidly targets the membrane fraction of macrophages and that it induces the formation of a plasma membrane pore. In vitro, the N‐terminal fragment of caspase‐1‐cleaved recombinant GSDMD tightly binds liposomes and forms large permeability pores. Visualization of liposome‐inserted GSDMD at nanometer resolution by cryo‐electron and atomic force microscopy shows circular pores with variable ring diameters around 20 nm. Overall, these data demonstrate that GSDMD is the direct and final executor of pyroptotic cell death.

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Section: Discussionsupporting

confidence: 79%

GSDMD membrane pore formation constitutes the mechanism of pyroptotic cell death

et al. 2016

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“…GSDMD 1-275 cleavage by caspase-1 induces pyroptosis by forming a large plasma membrane pore (28)(29)(30)(31)(32)(33). To evaluate the effect of EV71 3C on GSDMD, we assessed the cell viability and cell death of different cleavage fragments of GSDMD.…”

Section: Resultsmentioning

confidence: 99%

“…Upon activation by caspases, GSDMD undergoes proteolytic cleavage and releases its N-terminal domain, GSDMD 1-275 (28)(29)(30)(31)(32). As a result, this induces pyroptosis by causing extensive pore formation on the cell plasma membrane (28, 29, 31-33, 40, 41).…”

Section: Discussionmentioning

confidence: 99%

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Enterovirus 71 Inhibits Pyroptosis through Cleavage of Gasdermin D

Lei

Zhang

Xiao

et al. 2017

J Virol

120

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Enterovirus 71 (EV71) can cause hand-foot-and-mouth disease (HFMD) in young children. Severe infection with EV71 can lead to neurological complications and even death. However, the molecular basis of viral pathogenesis remains poorly understood. Here, we report that EV71 induces degradation of gasdermin D (GSDMD), an essential component of pyroptosis. Remarkably, the viral protease 3C directly targets GSDMD and induces its cleavage, which is dependent on the protease activity. Further analyses show that the Q193-G194 pair within GSDMD is the cleavage site of 3C. This cleavage produces a shorter N-terminal fragment spanning amino acids 1 to 193 (GSDMD ). However, unlike the N-terminal fragment produced by caspase-1 cleavage, this fragment fails to trigger cell death or inhibit EV71 replication. Importantly, a T239D or F240D substitution abrogates the activity of GSDMD consisting of amino acids 1 to 275 (GSDMD ). This is correlated with the lack of pyroptosis or inhibition of viral replication. These results reveal a previously unrecognized strategy for EV71 to evade the antiviral response.IMPORTANCE Recently, it has been reported that GSDMD plays a critical role in regulating lipopolysaccharide and NLRP3-mediated interleukin-1␤ (IL-1␤) secretion. In this process, the N-terminal domain of p30 released from GSDMD acts as an effector in cell pyroptosis. We show that EV71 infection downregulates GSDMD. EV71 3C cleaves GSDMD at the Q193-G194 pair, resulting in a truncated N-terminal fragment disrupted for inducing cell pyroptosis. Notably, GSDMD 1-275 (p30) inhibits EV71 replication whereas GSDMD 1-193 does not. These results reveal a new strategy for EV71 to evade the antiviral response. KEYWORDS EV71, HFMD, 3C, GSDMDE nterovirus 71 (EV71) is a major etiological agent of hand-foot-and-mouth disease (HFMD) which affects infants and children younger than 5 years of age. Most of these infections are self-limited with mild symptoms, including fever and vesicular eruptions mainly on the skin of hands and feet and in the mouth. However, some patients show severe neurological diseases, including aseptic meningitis, acute flaccid paralysis, encephalitis, and pulmonary edema. Since it was first isolated in California in 1974 (1-3), EV71 infection has caused many large-scale epidemics in the world, especially in the Asia-Pacific region (1, 4-6). To date, there are no effective therapeutic drugs for EV71 infection.EV71 belongs to the Enterovirus genus of the family Picornaviridae. The viral genome is approximately 7,500 nucleotides in length, with a single open reading frame that encodes a large precursor protein that is subsequently processed into three structural (VP0, VP1, and VP3) and seven nonstructural (2A, 2B, 2C, 3A, 3B, 3C, and 3D) proteins by the virus-encoded protease 2A or 3C (7). The three structural proteins and the RNA form provirions, in which VP0 is cleaved into VP2 and VP4 by RNA or 3CD, but the

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“…Independently of cytokine maturation, caspase-1 is able to cleave gasdermin D [44,45]. The N-terminal segment of gasdermin D forms pores in membranes [46][47][48][49] and induces a type of cell death called pyroptosis. In contrast to the pyrin domainefficient activation of Dronc, which dissociates and cleaves its substrate Drice (Fig.…”

Section: The Role Of Asc In Cytokine Maturation and Pyroptosismentioning

confidence: 99%

The NLRC4 inflammasome: The pieces of the puzzle are falling into place

Hafner-Bratkovič¹

2017

Inflammasome

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Inflammasomes are intracellular multiprotein platforms for the activation of inflammatory caspases. As components of the innate immune system, they play an important role in the fight against microbes. However, aberrant inflammasome activation has been implicated in auto-inflammatory syndromes. This review focuses on the NLRC4 inflammasome. This is perhaps not the most extensively studied, yet its mechanism of activation is by far the best understood. The NLRC4 inflammasome is activated by several proteins originating from intracellular bacteria, which are first sensed by receptors of the NAIP family. Activated NAIP binds NLRC4, which further recruits dormant NLRC4 molecules in a prion-like oligomerization event. NLRC4 enables a strong amplification of the signal, providing a fast and robust host response. The review also discusses peculiar NLRC4 inflammasome functions in promoting eicosanoid biosynthesis, actin reorganization, and its roles in autoinflammatory syndromes and sterile inflammation. Finally, the first inflammasomeindependent engagement of NLRC4 in suppressing melanoma tumor growth is presented. The emerging roles of NLRC4 in various normal and pathological processes demonstrate that there is still plenty to be learned about the NLRC4 mechanism of activation and downstream functions.

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GsdmD p30 elicited by caspase-11 during pyroptosis forms pores in membranes

Cited by 759 publications

References 27 publications

GSDMD membrane pore formation constitutes the mechanism of pyroptotic cell death

GSDMD membrane pore formation constitutes the mechanism of pyroptotic cell death

Enterovirus 71 Inhibits Pyroptosis through Cleavage of Gasdermin D

The NLRC4 inflammasome: The pieces of the puzzle are falling into place

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