title: Regulation of poly(ADP-ribose) polymerase-1-dependent gene expression through promoter-directed recruitment of a nuclear NAD+ synthase.
abstract: NMNAT-1 and PARP-1, two key enzymes in the NAD(+) metabolic pathway, localize to the nucleus where integration of their enzymatic activities has the potential to control a variety of nuclear processes. Using a variety of biochemical, molecular, cell-based, and genomic assays, we show that NMNAT-1 and PARP-1 physically and functionally interact at target gene promoters in MCF-7 cells. Specifically, we show that PARP-1 recruits NMNAT-1 to promoters where it produces NAD(+) to support PARP-1 catalytic activity, but also enhances the enzymatic activity of PARP-1 independently of NAD(+) production. Furthermore, using two-photon excitation microscopy, we show that NMNAT-1 catalyzes the production of NAD(+) in a nuclear pool that may be distinct from other cellular compartments. In expression microarray experiments, depletion of NMNAT-1 or PARP-1 alters the expression of about 200 protein-coding genes each, with about 10% overlap between the two gene sets. NMNAT-1 enzymatic activity is required for PARP-1-dependent poly(ADP-ribosyl)ation at the promoters of commonly regulated target genes, as well as the expression of those target genes. Collectively, our studies link the enzymatic activities of NMNAT-1 and PARP-1 to the regulation of a set of common target genes through functional interactions at target gene promoters.
authors.affiliation: Department of Molecular Biology and Genetics, Cornell University, Ithaca, New York 14853, USA.
authors.forename: Tong
authors.initials: T
authors.lastname: Zhang
authors.forename: Jhoanna G
authors.initials: JG
authors.lastname: Berrocal
authors.forename: Jie
authors.initials: J
authors.lastname: Yao
authors.forename: Michelle E
authors.initials: ME
authors.lastname: DuMond
authors.forename: Raga
authors.initials: R
authors.lastname: Krishnakumar
authors.forename: Donald D
authors.initials: DD
authors.lastname: Ruhl
authors.forename: Keun Woo
authors.initials: KW
authors.lastname: Ryu
authors.forename: Matthew J
authors.initials: MJ
authors.lastname: Gamble
authors.forename: W Lee
authors.initials: WL
authors.lastname: Kraus
mesh_terms: D021581:Active Transport, Cell Nucleus; D002460:Cell Line; D004789:Enzyme Activation; D020869:Gene Expression Profiling; D005786:Gene Expression Regulation; D006801:Humans; D009243:NAD / Q000378:metabolism; D009612:Nicotinamide-Nucleotide Adenylyltransferase / Q000378:metabolism / Q000502:physiology*; D000071137:Poly (ADP-Ribose) Polymerase-1; D011064:Poly Adenosine Diphosphate Ribose / Q000378:metabolism; D011065:Poly(ADP-ribose) Polymerases / Q000378:metabolism / Q000502:physiology*; D011401:Promoter Regions, Genetic; D011485:Protein Binding; D011499:Protein Processing, Post-Translational; D011506:Proteins / Q000378:metabolism; D060888:Real-Time Polymerase Chain Reaction; D014158:Transcription, Genetic
chemical_list: D011506:Proteins; C024857:poly(ADP)-ribosylated proteins; D009243:NAD; D011064:Poly Adenosine Diphosphate Ribose; C499320:PARP1 protein, human; D000071137:Poly (ADP-Ribose) Polymerase-1; D011065:Poly(ADP-ribose) Polymerases; C484007:NMNAT1 protein, human; D009612:Nicotinamide-Nucleotide Adenylyltransferase