Proline scanning mutagenesis reveals a role for the flap endonuclease-1 helical cap in substrate unpairing.

ABSTRACT: The prototypical 5'-nuclease, flap endonuclease-1 (FEN1), catalyzes the essential removal of single-stranded flaps during DNA replication and repair. FEN1 hydrolyzes a specific phosphodiester bond one nucleotide into double-stranded DNA. This specificity arises from double nucleotide unpairing that places the scissile phosphate diester on active site divalent metal ions. Also related to FEN1 specificity is the helical arch, through which 5'-flaps, but not continuous DNAs, can thread. The arch contains basic residues (Lys-93 and Arg-100 in human FEN1 (hFEN1)) that are conserved by all 5'-nucleases and a cap region only present in enzymes that process DNAs with 5' termini. Proline mutations (L97P, L111P, L130P) were introduced into the hFEN1 helical arch. Each mutation was severely detriment

SUBMITTER: Patel N 

PROVIDER: S-EPMC3837164 | biostudies-literature | 2013 Nov

REPOSITORIES: biostudies-literature