Isolation and expression of a human ornithine decarboxylase gene.
We have isolated a human ornithine decarboxylase (ODC) gene from a leukocyte genomic DNA library in order to examine the mechanisms involved in the regulation of ODC gene expression in normal and neoplastic cell growth. Nucleotide sequence analysis shows that the human ODC gene in clone ODC709-A2 consists of 12 exons which encode a protein identical to that inferred from a human ODC cDNA sequence. The 5' end of the gene was determined by S1 nuclease and primer extension mapping. The high G + C content and small open reading frame found in exon 1 may be pertinent to translation regulation of ODC. Conserved sequences and potential promoter elements including a TATA box, a possible CCAAT element, SP1 and AP-2 transcription factor binding sites, and cAMP response elements were identified in the 5'-flanking region. Transfection of mouse LM (tk-) cells with ODC709-A2 DNA resulted in the production of human ODC mRNA approximately 2.25 kilobases in length. Evidence that the protein synthesized from the human gene is functional is provided by "rescue" transfection of a Chinese hamster ovary mutant cell line, C55.7, which is ODC-deficient. C55.7 cells transfected with ODC709-A2 DNA expressed ODC enzyme activity and proliferated without exogenous putrescine.
Moshier
J A
JA
Department of Internal Medicine, Wayne State University School of Medicine, Detroit, Michigan 48201.
Gilbert
J D
JD
Skunca
M
M
Dosescu
J
J
Almodovar
K M
KM
Luk
G D
GD
D000818:Animals; D001483:Base Sequence; D002460:Cell Line; D003106:Colon / Q000201:enzymology; D003360:Cosmids; D004247:DNA / Q000235:genetics* / Q000302:isolation & purification; D015870:Gene Expression; D015723:Gene Library; D005796:Genes; D006801:Humans; D007413:Intestinal Mucosa / Q000201:enzymology*; D007739:L Cells / Q000201:enzymology; D051379:Mice; D008969:Molecular Sequence Data; D009693:Nucleic Acid Hybridization; D009955:Ornithine Decarboxylase / Q000235:genetics*; D011401:Promoter Regions, Genetic; D051381:Rats; D015183:Restriction Mapping; D012689:Sequence Homology, Nucleic Acid; D014162:Transfection
D004247:DNA; D009955:Ornithine Decarboxylase