Isolation and characterization of a cDNA clone that codes for human spermidine/spermine N1-acetyltransferase.
Spermidine/spermine N1-acetyltransferase (Spd/Spm acetyltransferase) is the rate-limiting enzyme in the catabolism of polyamines. This enzyme is highly inducible by several stimuli, including the natural polyamines and their structural analogues. To investigate the underlying mechanism responsible for the control of this enzyme a cDNA which codes for an active human Spd/Spm acetyltransferase has been isolated from a random primed cDNA library constructed from mRNA of a polyamine analogue treated large cell lung carcinoma line, NCI H157. The 972-base pair cDNA was identified using a 32-fold degenerate, 20-base oligomer probe to a 7-amino acid polypeptide sequence derived from the purified protein. The cDNA has a 513-base open reading frame that codes for a protein of 171 amino acids with a predicted molecular weight of 20,023. In vitro translation studies demonstrated the protein product of this cDNA to be a biologically active enzyme. The cDNA recognizes a 1.5-kilobase transcript in human cells which is highly induced in the human large cell lung carcinoma NCI H157 line following treatment with the polyamine analogue. The unusually high expression of Spd/Spm acetyltransferase mRNA by the NCI H157 cells in response to treatment does not appear to be a result of an amplification of the Spd/Spm acetyltransferase gene.
Casero
R A
RA
Johns Hopkins Oncology Center Laboratories, The Johns Hopkins School of Medicine, Baltimore, Maryland 21231.
Celano
P
P
Ervin
S J
SJ
Applegren
N B
NB
Wiest
L
L
Pegg
A E
AE
D000123:Acetyltransferases / Q000235:genetics*; D000595:Amino Acid Sequence; D001483:Base Sequence; D015152:Blotting, Northern; D015139:Blotting, Southern; D002999:Clone Cells; D004247:DNA / Q000737:chemistry / Q000302:isolation & purification*; D006801:Humans; D008969:Molecular Sequence Data; D014176:Protein Biosynthesis; D012333:RNA, Messenger / Q000235:genetics; D014407:Tumor Cells, Cultured; D000099228:Diamine N-Acetyltransferase
D000123:Acetyltransferases; D004247:DNA; D012333:RNA, Messenger; D000099228:Diamine N-Acetyltransferase