Cloning of rodent cDNA coding the poly(ADP-ribose) polymerase catalytic domain and analysis of mRNA levels during the cell cycle.
We have isolated a partial 2.0 kb cDNA (pRATC) encoding the entire 489 amino acids of the NAD binding domain located at the C terminus of the rat poly(ADP-ribose) polymerase. pRATC sequences were analysed and compared with the human mRNA. Our analysis reveals a remarkable homology between the rat and human nucleotide and amino acid sequences. Although a few minor amino acid changes were detected, we have found that the total number of possible phosphorylation sites remained constant in the NAD binding domain of both enzymes. We have also found that a 102 amino acid sequence, containing the putative nucleotide binding site Gly-Lys-Gly (position 378), is perfectly conserved between the rat and human sequences. Strong homology was also detected between pRATC and genomic DNA isolated from various vertebrates. In addition, we have analysed the levels of poly(ADP-ribose) polymerase mRNA throughout the cell cycle. Our results show that the levels of mRNA culminate in the G1 phase. We have also found that the increase in enzymatic activity observed in rats following treatment with phenobarbital did not correspond to an increase in the mRNA levels.
Thibodeau
J
J
Laboratoire du métabolisme du poly, Centre Hospitalier de l'Université Laval, Sainte-Foy, Québec, Canada.
Gradwohl
G
G
Dumas
C
C
Clairoux-Moreau
S
S
Brunet
G
G
Penning
C
C
Poirier
G G
GG
Moreau
P
P
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D001665:Binding Sites; D002384:Catalysis; D002453:Cell Cycle / Q000235:genetics; D002460:Cell Line; D003001:Cloning, Molecular; D004247:DNA / Q000235:genetics*; D015342:DNA Probes; D015723:Gene Library; D006801:Humans; D015151:Immunoblotting; D008969:Molecular Sequence Data; D010634:Phenobarbital / Q000494:pharmacology; D011065:Poly(ADP-ribose) Polymerases / Q000235:genetics*; D012333:RNA, Messenger / Q000378:metabolism*; D051381:Rats; D011919:Rats, Inbred Strains; D013045:Species Specificity
D015342:DNA Probes; D012333:RNA, Messenger; D004247:DNA; D011065:Poly(ADP-ribose) Polymerases; D010634:Phenobarbital