Visualization of poly(ADP-ribose) bound to PARG reveals inherent balance between exo- and endo-glycohydrolase activities

Eva Barkauskaite; Amy Brassington; Edwin S. Tan; Jim Warwicker; Mark S. Dunstan; Benito Banos; Pierre Lafite; Marijan Ahel; Timothy J. Mitchison; Ivan Ahel; David Leys

doi:10.1038/ncomms3164

Visualization of poly(ADP-ribose) bound to PARG reveals inherent balance between exo- and endo-glycohydrolase activities

Eva Barkauskaite, Amy Brassington, Edwin S. Tan, Jim Warwicker, Mark S. Dunstan, Benito Banos, Pierre Lafite, Marijan Ahel, Timothy J. Mitchison, Ivan Ahel, David Leys

Cancer Research UK Manchester Institute

Research output: Contribution to journal › Article › peer-review

Abstract

Poly-ADP-ribosylation is a post-translational modification that regulates processes involved in genome stability. Breakdown of the poly(ADP-ribose) (PAR) polymer is catalysed by poly(ADP-ribose) glycohydrolase (PARG), whose endo-glycohydrolase activity generates PAR fragments. Here we present the crystal structure of PARG incorporating the PAR substrate. The two terminal ADP-ribose units of the polymeric substrate are bound in exo-mode. Biochemical and modelling studies reveal that PARG acts predominantly as an exo-glycohydrolase. This preference is linked to Phe902 (human numbering), which is responsible for low-affinity binding of the substrate in endo-mode. Our data reveal the mechanism of poly-ADP-ribosylation reversal, with ADP-ribose as the dominant product, and suggest that the release of apoptotic PAR fragments occurs at unusual PAR/PARG ratios. © 2013 Macmillan Publishers Limited. All rights reserved.

Original language	English
Article number	2164
Journal	Nature Communications
Volume	4
DOIs	https://doi.org/10.1038/ncomms3164
Publication status	Published - 2013

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@article{65d24f7377d64fe192107f8b705bf771,

title = "Visualization of poly(ADP-ribose) bound to PARG reveals inherent balance between exo- and endo-glycohydrolase activities",

abstract = "Poly-ADP-ribosylation is a post-translational modification that regulates processes involved in genome stability. Breakdown of the poly(ADP-ribose) (PAR) polymer is catalysed by poly(ADP-ribose) glycohydrolase (PARG), whose endo-glycohydrolase activity generates PAR fragments. Here we present the crystal structure of PARG incorporating the PAR substrate. The two terminal ADP-ribose units of the polymeric substrate are bound in exo-mode. Biochemical and modelling studies reveal that PARG acts predominantly as an exo-glycohydrolase. This preference is linked to Phe902 (human numbering), which is responsible for low-affinity binding of the substrate in endo-mode. Our data reveal the mechanism of poly-ADP-ribosylation reversal, with ADP-ribose as the dominant product, and suggest that the release of apoptotic PAR fragments occurs at unusual PAR/PARG ratios. {\textcopyright} 2013 Macmillan Publishers Limited. All rights reserved.",

author = "Eva Barkauskaite and Amy Brassington and Tan, {Edwin S.} and Jim Warwicker and Dunstan, {Mark S.} and Benito Banos and Pierre Lafite and Marijan Ahel and Mitchison, {Timothy J.} and Ivan Ahel and David Leys",

year = "2013",

doi = "10.1038/ncomms3164",

language = "English",

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journal = "Nature Communications",

issn = "2041-1723",

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TY - JOUR

T1 - Visualization of poly(ADP-ribose) bound to PARG reveals inherent balance between exo- and endo-glycohydrolase activities

AU - Barkauskaite, Eva

AU - Brassington, Amy

AU - Tan, Edwin S.

AU - Warwicker, Jim

AU - Dunstan, Mark S.

AU - Banos, Benito

AU - Lafite, Pierre

AU - Ahel, Marijan

AU - Mitchison, Timothy J.

AU - Ahel, Ivan

AU - Leys, David

PY - 2013

Y1 - 2013

N2 - Poly-ADP-ribosylation is a post-translational modification that regulates processes involved in genome stability. Breakdown of the poly(ADP-ribose) (PAR) polymer is catalysed by poly(ADP-ribose) glycohydrolase (PARG), whose endo-glycohydrolase activity generates PAR fragments. Here we present the crystal structure of PARG incorporating the PAR substrate. The two terminal ADP-ribose units of the polymeric substrate are bound in exo-mode. Biochemical and modelling studies reveal that PARG acts predominantly as an exo-glycohydrolase. This preference is linked to Phe902 (human numbering), which is responsible for low-affinity binding of the substrate in endo-mode. Our data reveal the mechanism of poly-ADP-ribosylation reversal, with ADP-ribose as the dominant product, and suggest that the release of apoptotic PAR fragments occurs at unusual PAR/PARG ratios. © 2013 Macmillan Publishers Limited. All rights reserved.

AB - Poly-ADP-ribosylation is a post-translational modification that regulates processes involved in genome stability. Breakdown of the poly(ADP-ribose) (PAR) polymer is catalysed by poly(ADP-ribose) glycohydrolase (PARG), whose endo-glycohydrolase activity generates PAR fragments. Here we present the crystal structure of PARG incorporating the PAR substrate. The two terminal ADP-ribose units of the polymeric substrate are bound in exo-mode. Biochemical and modelling studies reveal that PARG acts predominantly as an exo-glycohydrolase. This preference is linked to Phe902 (human numbering), which is responsible for low-affinity binding of the substrate in endo-mode. Our data reveal the mechanism of poly-ADP-ribosylation reversal, with ADP-ribose as the dominant product, and suggest that the release of apoptotic PAR fragments occurs at unusual PAR/PARG ratios. © 2013 Macmillan Publishers Limited. All rights reserved.

U2 - 10.1038/ncomms3164

DO - 10.1038/ncomms3164

M3 - Article

C2 - 23917065

VL - 4

JO - Nature Communications

JF - Nature Communications

SN - 2041-1723

M1 - 2164

ER -

Visualization of poly(ADP-ribose) bound to PARG reveals inherent balance between exo- and endo-glycohydrolase activities

Abstract

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