Implications of the zinc-finger motif found in the DNA-binding domain of the human XPA protein.
BACKGROUND
The XPA (xeroderma pigmentosum group A) protein specifically recognizes the UV-or chemically damaged DNA lesions, and triggers the nucleotide excision repair process. This XPA protein contains the functional domain which is crucial to the recognition of damaged DNA. Its primary structure suggests that this DNA binding domain may contain a zinc-finger motif. To gain a more detailed insight into this zinc-finger motif, we have measured the 113Cd-NMR spectra of the DNA binding domains derived from the wild-type and mutant XPA proteins.


RESULTS
113Cd-NMR analysis, combined with atomic absorption and site-directed mutagenesis, revealed that the DNA binding domain contains one zinc ion, coordinated with four cysteine residues (Cys105, Cys108, Cys126, and Cys129), that is essential for correct protein folding in vivo and in vitro.


CONCLUSIONS
The four ligand cysteine residues form a Cys-X2-Cys-X17-Cys-X2-Cys motif, which is reminiscent of the (Cys)4 type zinc-finger motif found in numerous transcriptional regulatory proteins. However, the secondary structure prediction and the 3D-1D compatibility analysis demonstrate that there is no structural similarity in the vicinity of the zinc-finger motif between the XPA protein and other zinc-finger containing proteins. We conclude that the XPA protein contains a new type of zinc-finger motif.
Morita
E H
EH
Biomolecular Engineering Research Institute, Osaka, Japan.
Ohkubo
T
T
Kuraoka
I
I
Shirakawa
M
M
Tanaka
K
K
Morikawa
K
K
D000595:Amino Acid Sequence; D001665:Binding Sites / Q000235:genetics; D004260:DNA Repair; D004268:DNA-Binding Proteins / Q000737:chemistry* / Q000235:genetics*; D004926:Escherichia coli / Q000235:genetics; D006801:Humans; D008024:Ligands; D009682:Magnetic Resonance Spectroscopy; D008969:Molecular Sequence Data; D015394:Molecular Structure; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics; D013054:Spectrophotometry, Atomic; D051760:Xeroderma Pigmentosum Group A Protein; D015032:Zinc / Q000737:chemistry; D016335:Zinc Fingers / Q000235:genetics*
D004268:DNA-Binding Proteins; D008024:Ligands; D011994:Recombinant Proteins; C497314:XPA protein, human; D051760:Xeroderma Pigmentosum Group A Protein; D015032:Zinc