Differential splicing of the apoptosis-associated speck like protein containing a caspase recruitment domain (ASC) regulates inflammasomes.
Differential splicing of the apoptosis-associated speck like protein containing a caspase recruitment domain (ASC) regulates inflammasomes.
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DOI：
10.1186/1476-9255-7-23
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发表时间：
2010-05-18
期刊：
影响因子：
--
通讯作者：
Stehlik C
中科院分区：
文献类型：
--
作者：
Bryan NB;Dorfleutner A;Kramer SJ;Yun C;Rojanasakul Y;Stehlik C
凋亡斑点样蛋白（apoptosis speck-like protein，apoptosis sp它将激活的Nod样受体（NLR）（其是先天免疫系统的胞质模式识别受体家族）与半胱天冬酶1桥接，导致半胱天冬酶1激活和半胱天冬酶1底物的后续加工。因此，来自ASC缺陷型小鼠的巨噬细胞产生生物活性IL-1β的能力受损。此外，我们最近发现ASC响应于炎症刺激从细胞核易位到细胞溶质，以促进炎性体反应，其触发IL-1β加工和分泌。然而，在ASC水平上炎性小体的精确调节仍然没有完全理解。在这项研究中，我们确定并表征了三种新的ASC亚型作为炎性小体适配器的能力。为了确定ASC和ASC同种型作为功能性炎性体衔接子的能力，通过在炎性体重建测定中的ELISA、在THP-1和J774 A1细胞中的稳定表达以及通过恢复小鼠RAW 264.7巨噬细胞中内源性ASC的缺乏来研究IL-1β的加工和分泌。此外，通过免疫荧光染色确定ASC和ASC亚型的定位。三种新的ASC同种型ASC-b、ASC-c和ASC-d在其作为炎性体衔接子的功能方面显示出彼此和全长ASC独特且不同的能力，其中一种同种型甚至显示出抑制作用。一致的是，只有ASC的活化亚型ASC和ASC-b与NLRP 3和半胱天冬酶1共定位，而抑制亚型ASC-c仅与半胱天冬酶1共定位，而不与NLRP 3共定位。ASC-d不与NLRP 3或半胱天冬酶1共定位，并且始终缺乏作为炎性小体适配器发挥作用的能力，其精确功能和与ASC的关系需要进一步研究。选择性剪接和潜在的其他编辑机制产生ASC同种型，其具有作为炎性体衔接子的独特能力，其潜在地用于在炎性宿主应答期间调节炎性体。
The apoptotic speck-like protein containing a caspase recruitment domain (ASC) is the essential adaptor protein for caspase 1 mediated interleukin (IL)-1β and IL-18 processing in inflammasomes. It bridges activated Nod like receptors (NLRs), which are a family of cytosolic pattern recognition receptors of the innate immune system, with caspase 1, resulting in caspase 1 activation and subsequent processing of caspase 1 substrates. Hence, macrophages from ASC deficient mice are impaired in their ability to produce bioactive IL-1β. Furthermore, we recently showed that ASC translocates from the nucleus to the cytosol in response to inflammatory stimulation in order to promote an inflammasome response, which triggers IL-1β processing and secretion. However, the precise regulation of inflammasomes at the level of ASC is still not completely understood. In this study we identified and characterized three novel ASC isoforms for their ability to function as an inflammasome adaptor. To establish the ability of ASC and ASC isoforms as functional inflammasome adaptors, IL-1β processing and secretion was investigated by ELISA in inflammasome reconstitution assays, stable expression in THP-1 and J774A1 cells, and by restoring the lack of endogenous ASC in mouse RAW264.7 macrophages. In addition, the localization of ASC and ASC isoforms was determined by immunofluorescence staining. The three novel ASC isoforms, ASC-b, ASC-c and ASC-d display unique and distinct capabilities to each other and to full length ASC in respect to their function as an inflammasome adaptor, with one of the isoforms even showing an inhibitory effect. Consistently, only the activating isoforms of ASC, ASC and ASC-b, co-localized with NLRP3 and caspase 1, while the inhibitory isoform ASC-c, co-localized only with caspase 1, but not with NLRP3. ASC-d did not co-localize with NLRP3 or with caspase 1 and consistently lacked the ability to function as an inflammasome adaptor and its precise function and relation to ASC will need further investigation. Alternative splicing and potentially other editing mechanisms generate ASC isoforms with distinct abilities to function as inflammasome adaptor, which is potentially utilized to regulate inflammasomes during the inflammatory host response.
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DOI：
10.4049/jimmunol.0802367
发表时间：
2009-03-01
期刊：
Journal of immunology (Baltimore, Md. : 1950)
影响因子：
--
作者：
Bryan NB;Dorfleutner A;Rojanasakul Y;Stehlik C
通讯作者：
Stehlik C
DOI：
10.1073/pnas.0611496104
发表时间：
2007-05-08
影响因子：
11.1
作者：
Duncan, Joseph A.;Bergstralht, Daniel T.;Ting, Jenny Pan-Yun
通讯作者：
Ting, Jenny Pan-Yun
影响因子：
158.5
作者：
Jin, Ying;Mailloux, Christina M.;Spritz, Richard A.
通讯作者：
Spritz, Richard A.
影响因子：
30.5
作者：
Buerckstuemmer, Tilmann;Baumann, Christoph;Superti-Furga, Giulio
通讯作者：
Superti-Furga, Giulio
影响因子：
9.2
作者：
Feldmeyer, Laurence;Keller, Martin;Beer, Hans-Dietmar
通讯作者：
Beer, Hans-Dietmar