2. Academic Validation
  3. Chemical genetic discovery of PARP targets reveals a role for PARP-1 in transcription elongation

Chemical genetic discovery of PARP targets reveals a role for PARP-1 in transcription elongation

  • Science. 2016 Jul 1;353(6294):45-50. doi: 10.1126/science.aaf7865.
Bryan A Gibson 1,  Yajie Zhang 2,  Hong Jiang 3,  Kristine M Hussey 4,  Jonathan H Shrimp 3,  Hening Lin 3,  Frank Schwede 5,  Yonghao Yu 2,  W Lee Kraus 6
Affiliations

Affiliations

  • 1 The Laboratory of Signaling and Gene Expression, Cecil H. and Ida Green Center for Reproductive Biology Sciences and The Division of Basic Research, Department of Obstetrics and Gynecology, University of Texas Southwestern Medical Center, Dallas, TX 75390, USA.
  • 2 Department of Biochemistry, University of Texas Southwestern Medical Center, Dallas, TX 75390, USA.
  • 3 Howard Hughes Medical Institute and Department of Chemistry, Cornell University, Ithaca, NY 14850, USA.
  • 4 Sarepta Therapeutics, Cambridge, MA 02142, USA.
  • 5 Biolog Life Science Institute, D-28199 Bremen, Germany.
  • 6 The Laboratory of Signaling and Gene Expression, Cecil H. and Ida Green Center for Reproductive Biology Sciences and The Division of Basic Research, Department of Obstetrics and Gynecology, University of Texas Southwestern Medical Center, Dallas, TX 75390, USA. lee.kraus@utsouthwestern.edu.
PMID: 27256882 DOI: 10.1126/science.aaf7865
Abstract

Poly[adenosine diphosphate (ADP)-ribose] polymerases (PARPs) are a family of Enzymes that modulate diverse biological processes through covalent transfer of ADP-ribose from the oxidized form of nicotinamide adenine dinucleotide (NAD(+)) onto substrate proteins. Here we report a robust NAD(+) analog-sensitive approach for PARPs, which allows PARP-specific ADP-ribosylation of substrates that is suitable for subsequent copper-catalyzed azide-alkyne cycloaddition reactions. Using this approach, we mapped hundreds of sites of ADP-ribosylation for PARPs 1, 2, and 3 across the proteome, as well as thousands of PARP-1-mediated ADP-ribosylation sites across the genome. We found that PARP-1 ADP-ribosylates and inhibits negative elongation factor (NELF), a protein complex that regulates promoter-proximal pausing by RNA polymerase II (Pol II). Depletion or inhibition of PARP-1 or mutation of the ADP-ribosylation sites on NELF-E promotes Pol II pausing, providing a clear functional link between PARP-1, ADP-ribosylation, and NELF. This analog-sensitive approach should be broadly applicable across the PARP family and has the potential to illuminate the ADP-ribosylated proteome and the molecular mechanisms used by individual PARPs to mediate their responses to cellular signals.

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