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Molecular and Cellular Biology, January 2008, p. 814-824, Vol. 28, No. 2
0270-7306/08/$08.00+0     doi:10.1128/MCB.01766-07
Copyright © 2008, American Society for Microbiology. All Rights Reserved.

Functional Localization of Two Poly(ADP-Ribose)-Degrading Enzymes to the Mitochondrial Matrix{triangledown}

Marc Niere,1,{dagger} Stefan Kernstock,2,{dagger} Friedrich Koch-Nolte,2 and Mathias Ziegler1*

Department of Molecular Biology, University of Bergen, Thormøhlensgate 55, N-5008 Bergen, Norway,1 Institute of Immunology, University Hospital, Martinistrasse 52, D-20246 Hamburg, Germany2

Received 26 September 2007/ Returned for modification 25 October 2007/ Accepted 29 October 2007

Recent discoveries of NAD-mediated regulatory processes in mitochondria have documented important roles of this compartmentalized nucleotide pool in addition to energy transduction. Moreover, mitochondria respond to excessive nuclear NAD consumption arising from DNA damage-induced poly-ADP-ribosylation because poly(ADP-ribose) (PAR) can trigger the release of apoptosis-inducing factor from the organelles. To functionally assess mitochondrial NAD metabolism, we overexpressed the catalytic domain of nuclear PAR polymerase 1 (PARP1) and targeted it to the matrix, which resulted in the constitutive presence of PAR within the organelles. As a result, stably transfected HEK293 cells exhibited a decrease in NAD content and typical features of respiratory deficiency. Remarkably, inhibiting PARP activity revealed PAR degradation within mitochondria. Two enzymes, PAR glycohydrolase (PARG) and ADP-ribosylhydrolase 3 (ARH3), are known to cleave PAR. Both full-length ARH3 and a PARG isoform, which arises from alternative splicing, localized to the mitochondrial matrix. This conclusion was based on the direct demonstration of their PAR-degrading activity within mitochondria of living cells. The visualization of catalytic activity establishes a new approach to identify submitochondrial localization of proteins involved in the metabolism of NAD derivatives. In addition, targeted PARP expression may serve as a compartment-specific "knock-down" of the NAD content which is readily detectable by PAR formation.

* Corresponding author. Mailing address: Department of Molecular Biology, University of Bergen, Thormøhlensgate 55, N-5008 Bergen, Norway. Phone: 47 555 84591. Fax: 47 555 89683. E-mail: mathias.ziegler{at}mbi.uib.no

{triangledown} Published ahead of print on 8 November 2007.

{dagger} M.N. and S.K. contributed equally to this work.

Molecular and Cellular Biology, January 2008, p. 814-824, Vol. 28, No. 2
0270-7306/08/$08.00+0     doi:10.1128/MCB.01766-07
Copyright © 2008, American Society for Microbiology. All Rights Reserved.



This article has been cited by other articles:

  • Erdelyi, K., Bai, P., Kovacs, I., Szabo, E., Mocsar, G., Kakuk, A., Szabo, C., Gergely, P., Virag, L. (2009). Dual role of poly(ADP-ribose) glycohydrolase in the regulation of cell death in oxidatively stressed A549 cells. FASEB J. 23: 3553-3563 [Abstract] [Full Text]  
  • Buelow, B., Song, Y., Scharenberg, A. M. (2008). The Poly(ADP-ribose) Polymerase PARP-1 Is Required for Oxidative Stress-induced TRPM2 Activation in Lymphocytes. J. Biol. Chem. 283: 24571-24583 [Abstract] [Full Text]  


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