PARP-1 expression in the mouse is controlled by an autoregulatory loop: PARP-1 binding to an upstream S/MAR element and to a novel recognition motif in its promoter suppresses transcription

J Mol Biol. 2009 May 15;388(4):730-50. doi: 10.1016/j.jmb.2009.03.032. Epub 2009 Mar 18.

Abstract

This work identifies central components of a feedback mechanism for the expression of mouse poly(ADP-ribose) polymerase-1 (PARP-1). Using the stress-induced duplex destabilization algorithm, multiple base-unpairing regions (BURs) could be localized in the 5' region of the mouse PARP-1 gene (muPARP-1). Some of these could be identified as scaffold/matrix-attachment regions (S/MARs), suggesting an S/MAR-mediated transcriptional regulation. PARP-1 binding to the most proximal element, S/MAR 1, and to three consensus motifs, AGGCC, in its own promoter (basepairs -956 to +100), could be traced by electrophoretic mobility-shift assay. The AGGCC-complementary GGCCT motif was detected by cis-diammine-dichloro platinum cross-linking and functionally characterized by the effects of site-directed mutagenesis on its performance in wild type (PARP-1(+/+)) and PARP-1 knockout cells (PARP-1(-/-)). Mutation of the central AGGCC tract at basepairs -554 to -550 prevented PARP-1/promoter interactions, whereby muPARP-1 expression became up-regulated. Transfection of a series of reporter gene constructs with or without S/MAR 1 (basepairs -1523 to -1007) and the more distant S/MAR 2 (basepairs -8373 to -6880), into PARP-1(+/+) as well as PARP-1(-/-) cells, revealed an additional, major level of muPARP-1 promoter down-regulation, triggered by PARP-1 binding to S/MAR 1. We conclude that S/MAR 1 represents an upstream control element that acts in conjunction with the muPARP-1 promoter. These interactions are part of a negative autoregulatory loop.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Base Sequence
  • Binding Sites
  • Cells, Cultured
  • Cross-Linking Reagents / chemistry
  • DNA / chemistry
  • DNA / genetics
  • DNA / metabolism
  • DNA Damage
  • Fibroblasts / cytology
  • Fibroblasts / physiology
  • Formaldehyde / chemistry
  • Gene Expression Regulation*
  • Genes, Reporter
  • Humans
  • Mice
  • Mice, Knockout
  • Molecular Sequence Data
  • Poly (ADP-Ribose) Polymerase-1
  • Poly(ADP-ribose) Polymerases* / genetics
  • Poly(ADP-ribose) Polymerases* / metabolism
  • Promoter Regions, Genetic*
  • Protein Binding
  • Regulatory Sequences, Nucleic Acid*
  • Transcription, Genetic*

Substances

  • Cross-Linking Reagents
  • Formaldehyde
  • DNA
  • Parp1 protein, mouse
  • Poly (ADP-Ribose) Polymerase-1
  • Poly(ADP-ribose) Polymerases