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. 2009 Feb 12;457(7231):906-9.
doi: 10.1038/nature07575. Epub 2008 Dec 7.

ABIN-1 is a ubiquitin sensor that restricts cell death and sustains embryonic development

Shigeru Oshima  1 , Emre E Turer, Joseph A Callahan, Sophia Chai, Rommel Advincula, Julio Barrera, Nataliya Shifrin, Bettina Lee, T S Benedict Yen, Tammy Woo, Barbara A Malynn, Averil Ma
Affiliations

ABIN-1 is a ubiquitin sensor that restricts cell death and sustains embryonic development

Shigeru Oshima et al. Nature. .

Erratum in

  • Nature. 2009 Mar 26;458(7237):538

Abstract

Proteins that directly regulate tumour necrosis factor receptor (TNFR) signalling have critical roles in regulating cellular activation and survival. ABIN-1 (A20 binding and inhibitor of NF-kappaB) is a novel protein that is thought to inhibit NF-kappaB signalling. Here we show that mice deficient for ABIN-1 die during embryogenesis with fetal liver apoptosis, anaemia and hypoplasia. ABIN-1 deficient cells are hypersensitive to tumour necrosis factor (TNF)-induced programmed cell death, and TNF deficiency rescues ABIN-1 deficient embryos. ABIN-1 inhibits caspase 8 recruitment to FADD (Fas-associated death domain-containing protein) in TNF-induced signalling complexes, preventing caspase 8 cleavage and programmed cell death. Moreover, ABIN-1 directly binds polyubiquitin chains and this ubiquitin sensing activity is required for ABIN-1's anti-apoptotic activity. These studies provide insights into how ubiquitination and ubiquitin sensing proteins regulate cellular and organismal survival.

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Figures

Figure 1
Figure 1. ABIN-1 is required for embryonic development
(A) Gross appearance of ABIN-1+/+ and ABIN-1−/− embryos. (B) Hypoplasia of E18.5 ABIN-1−/− embryos. Weights of individual embryos are shown as circles; horizontal bars indicate mean weights for each genotype. ABIN-1−/− embryos weigh less than ABIN-1+/+ and ABIN-1+/− control embryos (p < 0.01 between ABIN-1+/+ and ABIN-1−/− embryos; and p < 0.01 between ABIN-1+/− and ABIN-1−/− embryos, n=35). (C) Anemia of E18.5 ABIN1−/− embryos. ABIN-1−/− embryos are markedly anemic (p < 0.02 between ABIN-1+/+ and ABIN-1−/− embryos; p < 0.01 between ABIN-1+/− and ABIN-1−/− embryos, n=18). (D) Hematoxilin and eosin (H&E) histology (upper panels) and cleaved caspase 3 immunohistochemistry (lower panels) of sequential sections from ABIN-1+/+ and ABIN-1−/− fetal livers. Apoptotic patches were found in 5 of 7 ABIN-1−/− fetal livers analyzed and in none of 10 ABIN-1+/− and ABIN+/+ fetal livers analyzed (p < 0.01).
Figure 2
Figure 2. ABIN-1 is required for protecting cells from TNF-induced PCD
(A) TNF-induced PCD of ABIN-1+/+ and ABIN-1−/− MEF (p < 0.01 between ABIN-1+/+ and ABIN-1−/− cells; means and standard deviations indicated, n=3). (B) TNF-induced caspase-3 and BID expression in ABIN-1+/+ (left) and ABIN-1−/− (right) MEFs. Cleaved, active forms of Bid (tBid) and caspase 3 indicated by arrows. Actin expression shown below as protein loading control. (C) TNF-induced PCD of ABIN-1−/− 3T3s expressing GFP and FLAG-ABIN-1 (black columns on right) or GFP alone (white columns on left), after treatment with the indicated agents. Means and standard deviations indicated, n=3. (D) TNF-induced caspase-8, caspase 3, and BID expression in ABIN-1−/− 3T3s expressing GFP and FLAG-ABIN-1 (left) or GFP alone (right). Cleaved, active forms of caspase 8, caspase 3 and Bid (tBid) indicated by arrows. Actin expression shown below as control. (E) TNF-induced expression of MAP kinase signaling proteins and A20 in ABIN-1−/− and control MEFs. (F) TNF-induced recruitment of RIP1 and FADD to Myc (ABIN-1). Expression of Myc-ABIN-1 in IPs, and expression of FADD and caspase 8 in whole cell lysates (“input”) shown below as controls. (G) TNF-induced recruitment of RIP1, FADD and A20 to caspase 8 in ABIN-1 deficient and control HT1080 cells. Expression of caspase 8 in IPs and expression of ABIN-1, FADD and actin in whole cell lysates (“input”) shown below as controls. All data are representative of 3–5 independent experiments.
Figure 3
Figure 3. ABIN-1 is a ubiquitin sensor that uses a NUB domain to bind to DISC signaling complexes and protect cells from TNF-induced PCD
(A) Sequence alignment of AHD2 (and neighboring sequences) of ABIN-1 with the NEMO ubiquitin binding (NUB) domain of NEMO/IKKγ. Thick bars mark locations of residues mutated in QQ477EE, F482S, and D485N ABIN-1 mutants. (B) Ubiquitin binding to GST-ABIN-1 in GST pull down assay. Ubiquitin bound to GST-ABIN-1 shown in left six lanes; input of ubiquitin chains is shown as a control in right three lanes. (C) GST-ABIN-1 proteins bound to K63-linked His-polyubiquitin chains in Nickel (Ni) agarose pull down assay. GST-ABIN-1 proteins bound to His-ubiquitin chains shown in upper panel; inputs of GST-ABIN-1 proteins shown in bottom panel as controls. (D) 6xHis-K63 linked ubiquitin chains bound to GST-C-terminal (wild type and mutant) ABIN-1 proteins. Input levels of GST-ABIN-1 proteins shown below as controls. (E) TNF-induced PCD of ABIN-1−/− 3T3s virally complemented with the indicated FLAG-ABIN-1 proteins or GFP alone. FLAG-ABIN-1 expression levels shown below as control. (F) Recruitment of RIP1 or FADD to wild type (WT) or mutant Myc-ABIN-1 proteins. Myc-ABIN-1 expression in anti-Myc immunoprecipitates and RIP1 and FADD expression in whole cell lysates (“input”) shown below as controls. (G) Recruitment of FADD to caspase 8 in presence of wild type, or QQ477EE mutant FLAG-ABIN-1 proteins, or no ABIN-1 proteins (“GFP”). Expression of caspase 8 in IPs and expression of FLAG-ABIN-1, caspase 8, FADD and actin in whole cell lysates (“input”) shown below as controls. (H) Recruitment of A20 to wild type or mutant (“QQ477EE”) FLAG-ABIN-1 proteins. Expression of FLAG-ABIN-1 in IPs and A20 in whole cell lysates (“input”) shown as controls. (I) Anti-apoptotic activity of ABIN-1 in A20−/− 3T3s expressing no ABIN-1 (“GFP”), WT ABIN-1 (“WT”), or mutant ABIN-1 (“QQ477EE”). Means and standard deviations indicated, n=3. Expression of FLAG-ABIN-1, A20, and actin proteins shown as controls at left (N.S. = non-specific band directly above FLAG-ABIN-1 band). All data are representative of 3–5 independent experiments.
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References

    1. Heyninck K, De Valck D, Vanden Berghe W, Van Criekinge W, Contreras R, Fiers W, Haegeman G, Beyaert R. The zinc finger protein A20 inhibits TNF-induced NF-kappaB-dependent gene expression by interfering with an RIP- or TRAF2-mediated transactivation signal and directly binds to a novel NF-kappaB-inhibiting protein ABIN-1. J Cell Biol. 1999;145:1471–1482. - PMC - PubMed
    1. Opipari AW, Jr, Boguski MS, Dixit VM. The A20 cDNA induced by tumor necrosis factor alpha encodes a novel type of zinc finger protein. J Biol Chem. 1990;265:14705–14708. - PubMed
    1. Doi TS, Marino MW, Takahashi T, Yoshida T, Sakakura T, Old LJ, Obata Y. Absence of tumor necrosis factor rescues RelA-deficient mice from embryonic lethality. Proc Natl Acad Sci U S A. 1999;96:2994–2999. - PMC - PubMed
    1. Beg AA, Baltimore D. An essential role for NF-kappaB in preventing TNF-alpha-induced cell death. Science. 1996;274:782–784. - PubMed
    1. Wang CY, Mayo MW, Baldwin AS., Jr TNF- and cancer therapy-induced apoptosis: potentiation by inhibition of NF-kappaB. Science. 1996;274:784–787. - PubMed

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