Mouse Rt6.1 is a thiol‐dependent arginine‐specific ADP‐ribosyltransferase

Hara, Nobumasa; Badruzzaman, Muhammad; Sugae, Takashi; Shimoyama, Makoto; Tsuchiya, Masaharu

doi:10.1046/j.1432-1327.1999.00039.x

Cited by 16 publications

(15 citation statements)

References 38 publications

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“…In mice, ART activity has also been detected on lymphoid cells, including T cell lymphomas (11,12), splenocytes (13,14), and activated CTL (15) and is believed to participate in the regulation of cell functions. A 35-kDa GPI-anchored ART was found to mediate inhibition of proliferation and cytotoxic activity of CTL cell lines in the presence of extracellular NAD (16).…”

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confidence: 99%

Rapid Induction of Naive T Cell Apoptosis by Ecto-Nicotinamide Adenine Dinucleotide: Requirement for Mono(ADP-Ribosyl)Transferase 2 and a Downstream Effector

Adriouch

Ohlrogge²,

Haag³

et al. 2001

The Journal of Immunology

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Lymphocytes express a number of NAD-metabolizing ectoenzymes, including mono(ADP-ribosyl)transferases (ART) and ADP ribosylcyclases. These enzymes may regulate lymphocyte functions following the release of NAD in injured or inflammatory tissues We report here that extracellular NAD induces apoptosis in BALB/c splenic T cells with an IC50 of 3–5 μM. Annexin V staining of cells was observed already 10 min after treatment with NAD in the absence of any additional signal. Removal of GPI-anchored cell surface proteins by phosphatidylinositol-specific phospholipase C treatment rendered cells resistant to NAD-mediated apoptosis. RT-PCR analyses revealed that resting BALB/c T cells expressed the genes for GPI-anchored ART2.1 and ART2.2 but not ART1. ART2-specific antisera blocked radiolabeling of cell surface proteins with both [32P]NAD and NAD-mediated apoptosis. Further analyses revealed that natural knockout mice for Art2.a (C57BL/6) or Art2.b (NZW) were resistant to NAD-mediated apoptosis. Labeling with [32P]NAD revealed strong cell surface ART activity on T cells of C57BL/6 and little if any activity on cells of NZW mice. T cells of (C57BL/6 × NZW)F1 animals showed strong cell surface ART activity and were very sensitive to NAD-induced apoptosis. As in BALB/c T cells, ART2-specific antisera blocked cell surface ART activity and apoptosis in (C57BL/6 × NZW)F1 T cells. The fact that T cells of F1 animals are sensitive to rapid NAD-induced apoptosis suggests that this effect requires the complementation of (at least) two genetic components. We propose that one of these is cell surface ART2.2 activity (defective in the NZW parent), the other a downstream effector of ADP-ribosylation (defective in the C57BL/6 parent).

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confidence: 99%

Rapid Induction of Naive T Cell Apoptosis by Ecto-Nicotinamide Adenine Dinucleotide: Requirement for Mono(ADP-Ribosyl)Transferase 2 and a Downstream Effector

Adriouch

Ohlrogge²,

Haag³

et al. 2001

The Journal of Immunology

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“…However, an additional pair of unique cysteines (Cys-80 and Cys-201) in ART2.1 can result in disulfide bond formation that allosterically suppresses catalytic activity of this isoform. This inhibited state of ART2.1 is readily reversed in the presence of thiol reductants, such as exogenous dithiothreitol or endogenous cysteine or glutathione, that accumulate in the extracellular compartments of inflamed or hypoxically stressed tissues [11,12]. The additional layer of allosteric regulation for ART2.1, but not ART2.2, indicates that these isoforms are not simply redundant gene products; this is also consistent with their differential expression in various inbred mouse strains [13][14][15].…”

Section: Introductionmentioning

confidence: 56%

Basal and inducible expression of the thiol-sensitive ART2.1 ecto-ADP-ribosyltransferase in myeloid and lymphoid leukocytes

Hong

Brass²,

Séman

et al. 2009

Purinergic Signalling

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ADP-ribosylation of cell surface proteins in mammalian cells is a post-translational modification by which ecto-ADP-ribosyltransferases (ARTs) transfer ADPribose from extracellular NAD to protein targets. The ART2 locus at murine chromosome 7 encompasses the tandem Art2a and Art2b genes that encode the distinct ART2.1 and ART2.2 proteins. Although both ecto-enzymes share 80% sequence identity, ART2.1 activity is uniquely regulated by an allosteric disulfide bond that is reducible in the presence of extracellular thiols, such as cysteine and glutathione, that accumulate in hypoxic and ischemic tissues. Previous studies have characterized the expression of ART2.1 and ART2.2 in murine T lymphocytes but not in other major classes of lymphoid and myeloid leukocytes. Here, we describe the expression of ART2.1 activity in a wide range of freshly isolated or tissue-cultured murine myeloid and lymphoid leukocytes. Spleen-derived macrophages, dendritic cells (DC), and B cells constitutively express ART2.1 as their predominant ART while spleen T cells express both ART2.1 and the thiol-independent ART2.2 isoform. Although bone-marrow-derived macrophages (BMDM) and dendritic cells (BMDC) constitutively express ART2.1 at low levels, it is markedly up-regulated when these cells are stimulated in vitro with IFNβ or IFNγ. ART2.1 expression and activity in splenic B cells is modestly up-regulated during incubation in vitro for 24 h, a condition that promotes B cell apoptosis. This increase in ART2.1 is attenuated by IL-4 (a B cell survival factor), but is not affected by IFNβ/γ, suggesting a possible induction of ART2.1 as an ancillary response to B cell apoptosis. In contrast, ART2.1 and ART2.2, which are highly expressed in freshly isolated splenic T cells, are markedly downregulated when purified T cells are incubated in vitro for 12-24 h. Studies with the BW5147 mouse thymocyte line verified basal expression of ART2.1 and ART2.2, as in primary spleen T cells, and demonstrated that both isoforms can be up-regulated when T cells are maintained in the presence of IFNs. Comparison of the surface proteins which are ADP-ribosylated by ART2.1 in the different leukocyte subtypes indicated both shared and cell-specific proteins as ART2.1 substrates. The LFA-1 integrin, a major target for ART2.2 in T cells, is also ADP-ribosylated by the ART2.1 expressed in macrophages. Thus, ART2.1, in contrast to ART2.2, is expressed in a broad range of myeloid and lymphoid leukocytes. The thiol redox-sensitive nature of this ecto-enzyme suggests an involvement in purinergic signaling that occurs in the combined context of inflammation and hypoxia/ischemia.

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“…5D). Although both ART2.1 and ART2.2 act as ecto-ARTs, only ART2.1 requires the presence of extracellular thiol cofactors to modulate a reactive cysteine uniquely present at position 201 in ART2.1 (19,20). Interestingly, in the three-dimensional structure of rat ART2.2, the side chains of Ser 60 and Phe 181 , i.e., the residues corresponding to the extra cysteine residues in ART2.1, are in close proximity, suggesting that the two cysteines may form a labile disulfide bridge in ART2.1 (Fig.…”

Section: Discussionmentioning

confidence: 99%

“…Both ART2.1 and ART2.2 can act as either ART or NAD-glycohydrolase. However, the activity of ART2.1 requires the presence of extracellular thiol-reducing agents, such as DTT or cysteine, to reversibly control oxidation of unique residues (Cys 80 and Cys 201 ) present in ART2.1 but not ART2.2 (19,20). In the presence of micromolar extracellular NAD, ART2 enzymes modulate a variety of murine T lymphocyte functions by ADP-ribosylating multiple cell surface proteins including CD8, CD44, CD45, and the CD11a and CD18 chains of LFA-1 (12,13).…”

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confidence: 99%

Lipopolysaccharide, IFN-γ, and IFN-β Induce Expression of the Thiol-Sensitive ART2.1 Ecto-ADP-Ribosyltransferase in Murine Macrophages

Hong

Brass²,

Séman

et al. 2007

The Journal of Immunology

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Extracellular NAD modulates multiple immune and inflammatory responses via the activity of a family of ecto‐ADP‐ribosyltransferases (ARTs) that covalently modify target proteins at arginine or cysteine residues. Functional ARTs have been identified thus far in neutrophils and T cells but not macrophages. This study investigates the expression and function of ARTs by RT‐PCR and western blot analysis in primary bone‐marrow‐derived macrophages (BMDM) isolated from Balb/c mice. Although naive BMDM did not express any of the 6 murine ART genes at the mRNA or protein level, stimulation with lipopolysaccharide (LPS) or ligands for other Toll‐like receptors induced robust expression of ART2.1 as a GPI‐anchored cell surface ecto‐enzyme. ART2.1 expression was potentiated by pharmacological inhibition of ERK1/2 pathways but inhibited by pharmacological blockade of NFkappaB‐ or PI3K‐ signaling pathways. Significantly, catalytic enzyme activity of the inducible ART2.1 was strictly dependent on the presence of extracellular thiol reducing molecules such as cysteine or dithiolthreitol. These data indicate that modulation of immune function by NAD‐dependent ART2.1 activity in macrophages may be synergistically regulated by extracellular redox environments that occur during hypoxia or ishchemia. Support: NIH‐GM‐36387

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Mouse Rt6.1 is a thiol‐dependent arginine‐specific ADP‐ribosyltransferase

Cited by 16 publications

References 38 publications

Rapid Induction of Naive T Cell Apoptosis by Ecto-Nicotinamide Adenine Dinucleotide: Requirement for Mono(ADP-Ribosyl)Transferase 2 and a Downstream Effector

Rapid Induction of Naive T Cell Apoptosis by Ecto-Nicotinamide Adenine Dinucleotide: Requirement for Mono(ADP-Ribosyl)Transferase 2 and a Downstream Effector

Basal and inducible expression of the thiol-sensitive ART2.1 ecto-ADP-ribosyltransferase in myeloid and lymphoid leukocytes

Lipopolysaccharide, IFN-γ, and IFN-β Induce Expression of the Thiol-Sensitive ART2.1 Ecto-ADP-Ribosyltransferase in Murine Macrophages

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