Synthesis of a new zinc finger peptide; comparison of its 'code' deduced and 'CASTing' derived binding sites.
Using two synthetic oligonucleotides, we have constructed a new gene containing three zinc finger motifs of the Cys2-His2 type. We named this artificial gene 'Mago'. The Mago nucleotide triplets encoding the amino acid positions, described to be crucial for DNA binding specificity, have been chosen on the basis of the proposed recognition 'code' that relates the zinc finger's primary structure to the DNA binding target. Here we demonstrate that Mago protein specifically binds the 'code' DNA target, with a dissociation constant (Kd) comparable to the Kd of the well known Zif268 protein with its binding site. Moreover, we show that the deduced Mago 'code' and the 'experimental' selected DNA binding sites are almost identical, differing only in two nucleotides at the side positions.
Corbi
N
N
Dipartimento di Biotecnologie Cellulari ed Ematologia, Università di Roma La Sapienza, Rome, Italy.
Perez
M
M
Maione
R
R
Passananti
C
C
D000595:Amino Acid Sequence; D001483:Base Sequence; D001665:Binding Sites; D004247:DNA / Q000378:metabolism; D004268:DNA-Binding Proteins / Q000138:chemical synthesis* / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D008969:Molecular Sequence Data; D010455:Peptides / Q000138:chemical synthesis* / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D015202:Protein Engineering; D016335:Zinc Fingers
D004268:DNA-Binding Proteins; C109502:Mago protein, synthetic; D010455:Peptides; D004247:DNA