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

• DOI: 10.1126/science.aaf7865
• Publication Date: 2016-06-03T03:33:41Z
• Authors: Bryan A. Gibson; Yajie Zhang; Hong Jiang; Kristine M. Hussey; Jonathan H. Shrimp; Hening Lin; Frank Schwede; Yonghao Yu; W. Lee Kraus
• Keywords: ADP-ribosylation; RNA polymerase II; Proteome

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