title: DNA binding specificities of the long zinc-finger recombination protein PRDM9.
abstract: BACKGROUND
  Meiotic recombination ensures proper segregation of homologous chromosomes and creates genetic variation. In many organisms, recombination occurs at limited sites, termed 'hotspots', whose positions in mammals are determined by PR domain member 9 (PRDM9), a long-array zinc-finger and chromatin-modifier protein. Determining the rules governing the DNA binding of PRDM9 is a major issue in understanding how it functions.
  
  
  RESULTS
  Mouse PRDM9 protein variants bind to hotspot DNA sequences in a manner that is specific for both PRDM9 and DNA haplotypes, and that in vitro binding parallels its in vivo biological activity. Examining four hotspots, three activated by Prdm9Cst and one activated by Prdm9Dom2, we found that all binding sites required the full array of 11 or 12 contiguous fingers, depending on the allele, and that there was little sequence similarity between the binding sites of the three Prdm9Cst activated hotspots. The binding specificity of each position in the Hlx1 binding site, activated by Prdm9Cst, was tested by mutating each nucleotide to its three alternatives. The 31 positions along the binding site varied considerably in the ability of alternative bases to support binding, which also implicates a role for additional binding to the DNA phosphate backbone.
  
  
  CONCLUSIONS
  These results, which provide the first detailed mapping of PRDM9 binding to DNA and, to our knowledge, the most detailed analysis yet of DNA binding by a long zinc-finger array, make clear that the binding specificities of PRDM9, and possibly other long-array zinc-finger proteins, are unusually complex.
authors.forename: Timothy
authors.initials: T
authors.lastname: Billings
authors.forename: Emil D
authors.initials: ED
authors.lastname: Parvanov
authors.forename: Christopher L
authors.initials: CL
authors.lastname: Baker
authors.forename: Michael
authors.initials: M
authors.lastname: Walker
authors.forename: Kenneth
authors.initials: K
authors.lastname: Paigen
authors.forename: Petko M
authors.initials: PM
authors.lastname: Petkov
mesh_terms: D000818:Animals; D001665:Binding Sites; D004247:DNA / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D011495:Histone-Lysine N-Methyltransferase / Q000235:genetics / Q000378:metabolism*; D051379:Mice; D008810:Mice, Inbred C57BL; D009154:Mutation; D059372:Nucleotide Motifs; D011485:Protein Binding
chemical_list: D004247:DNA; D011495:Histone-Lysine N-Methyltransferase; C534829:prdm9 protein, mouse