PARP14 is a writer, reader, and eraser of mono-ADP-ribosylation
- J Biol Chem. 2023 Sep;299(9):105096. doi: 10.1016/j.jbc.2023.105096.
- 1. Department of Chemistry, Center for Molecular Protein Science (CMPS), Lund University, Lund, Sweden.
- 2. Department of Chemistry, Center for Molecular Protein Science (CMPS), Lund University, Lund, Sweden. Electronic address: [email protected].
PARP14/BAL2 is a large multidomain enzyme involved in signaling pathways with relevance to Cancer, inflammation, and Infection. Inhibition of its mono-ADP-ribosylating PARP homology domain and its three ADP-ribosyl binding macro domains has been regarded as a potential means of therapeutic intervention. Macrodomains-2 and -3 are known to stably bind to ADP-ribosylated target proteins, but the function of macrodomain-1 has remained somewhat elusive. Here, we used biochemical assays of ADP-ribosylation levels to characterize PARP14 macrodomain-1 and the homologous macrodomain-1 of PARP9. Our results show that both macrodomains display an ADP-ribosyl glycohydrolase activity that is not directed toward specific protein side chains. PARP14 macrodomain-1 is unable to degrade poly(ADP-ribose), the enzymatic product of PARP1. The F926A mutation of PARP14 and the F244A mutation of PARP9 strongly reduced ADP-ribosyl glycohydrolase activity of the respective macrodomains, suggesting mechanistic homology to the Mac1 domain of the SARS-CoV-2 Nsp3 protein. This study adds two new Enzymes to the previously known six human ADP-ribosyl glycohydrolases. Our results have key implications for how PARP14 and PARP9 will be studied and how their functions will be understood.