GSBS Student Publications
Title
ART2, a T cell surface mono-ADP-ribosyltransferase, generates extracellular poly(ADP-ribose)
GSBS Program
Interdisciplinary Graduate Program
Publication Date
2006-11-03
UMMS Affiliation
Graduate School of Biomedical Sciences; Department of Biochemistry and Molecular Pharmacology; Department of Medicine; Senior Scholars Program
Document Type
Article
Disciplines
Biochemistry
Abstract
NAD functions in multiple aspects of cellular metabolism and signaling through enzymes that covalently transfer ADP-ribose from NAD to acceptor proteins, thereby altering their function. NAD is a substrate for two enzyme families, mono-ADP-ribosyltransferases (mARTs) and poly(ADP-ribose) polymerases (PARPs), that covalently transfer an ADP-ribose monomer or polymer, respectively, to acceptor proteins. ART2, a mART, is a phenotypic marker of immunoregulatory cells found on the surface of T lymphocytes, including intestinal intraepithelial lymphocytes (IELs). We have shown that the auto-ADP-ribosylation of the ART2.2 allelic protein is multimeric. Our backbone structural alignment of ART2 (two alleles of the rat art2 gene have been reported, for simplicity, the ART2.2 protein investigated in this study will be referred to as ART2) and PARP suggested that multimeric auto-ADP-ribosylation of ART2 may represent an ADP-ribose polymer, rather than multiple sites of mono-ADP-ribosylation. To investigate this, we used highly purified recombinant ART2 and demonstrated that ART2 catalyzes the formation of an ADP-ribose polymer by sequencing gel and by HPLC and MS/MS mass spectrometry identification of PR-AMP, a breakdown product specific to poly(ADP-ribose). Furthermore, we identified the site of ADP-ribose polymer attachment on ART2 as Arg-185, an arginine in a crucial loop of its catalytic core. We found that endogenous ART2 on IELs undergoes multimeric auto-ADP-ribosylation more efficiently than ART2 on peripheral T cells, suggesting that these distinct lymphocyte populations differ in their ART2 surface topology. Furthermore, ART2.2 IELs are more resistant to NAD-induced cell death than ART2.1 IELs that do not have multimeric auto-ADP-ribosylation activity. The data suggest that capability of polymerizing ADP-ribose may not be unique to PARPs and that poly(ADP-ribosylation), an established nuclear activity, may occur extracellularly and modulate cell function.
DOI of Published Version
10.1074/jbc.M607259200
Source
J Biol Chem. 2006 Nov 3;281(44):33363-72. Epub 2006 Aug 24. Link to article on publisher's site
Journal/Book/Conference Title
The Journal of biological chemistry
Related Resources
PubMed ID
16931513
Repository Citation
Morrison, Alan R.; Moss, Joel; Stevens, Linda A.; Evans, James E.; Farrell, Caitlin; Merithew, Eric Lee; Lambright, David G.; Greiner, Dale L.; Mordes, John P.; Rossini, Aldo A.; and Bortell, Rita, "ART2, a T cell surface mono-ADP-ribosyltransferase, generates extracellular poly(ADP-ribose)" (2006). GSBS Student Publications. 884.
https://escholarship.umassmed.edu/gsbs_sp/884
Comments
Medical student Alan Morrison participated in this study as part of the Senior Scholars research program.