Cloning of a full-length cDNA encoding bovine thymus poly(ADP-ribose) synthetase: evolutionarily conserved segments and their potential functions.
The primary structure of bovine thymus poly(ADP-ribose) synthetase, as deduced from the nucleotide sequence of a cloned cDNA, indicated that this enzyme is composed of 1016 amino acids (aa) with an Mr of 113,481. An abundance of Lys and Arg residues was in accord with the known basic nature of this protein. A comparison with reported sequences of human counterparts revealed: (1) three functional domains separated by partial proteolysis, i.e., DNA-binding (N-terminal), automodification (central), and NAD-binding (C-terminal) domains, have, in this order, increasing degrees of homology; (2) the DNA-binding domain is composed of two distinct regions: one, less conserved, containing zinc-binding fingers and the other, more conserved, containing helix-turn-helix motifs; (3) all Glu and Asp residues in the automodification domain are conserved; and (4) a 78-aa stretch encompassing the nucleotide-binding fold in the NAD-binding domain is completely conserved. These results are compatible with specific features of each domain, i.e., complex DNA-enzyme interactions, multiple automodification at acidic aa residues, and a stringent specificity for the substrate, NAD.
Saito
I
I
Department of Clinical Science, Kyoto University Faculty of Medicine, Japan.
Hatakeyama
K
K
Kido
T
T
Ohkubo
H
H
Nakanishi
S
S
Ueda
K
K
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D005075:Biological Evolution; D002417:Cattle; D003001:Cloning, Molecular; D004247:DNA / Q000096:biosynthesis*; D004268:DNA-Binding Proteins / Q000235:genetics / Q000378:metabolism; D015698:Genomic Library; D006801:Humans; D007231:Infant, Newborn; D008667:Metalloproteins / Q000235:genetics / Q000378:metabolism; D008969:Molecular Sequence Data; D009243:NAD / Q000378:metabolism; D011065:Poly(ADP-ribose) Polymerases / Q000096:biosynthesis / Q000235:genetics*; D015183:Restriction Mapping; D012689:Sequence Homology, Nucleic Acid; D013329:Structure-Activity Relationship; D013950:Thymus Gland / Q000201:enzymology*; D015032:Zinc / Q000378:metabolism
D004268:DNA-Binding Proteins; D008667:Metalloproteins; D009243:NAD; D004247:DNA; D011065:Poly(ADP-ribose) Polymerases; D015032:Zinc