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 co...

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Bibliographic Details
Published in:	The Journal of biological chemistry Vol. 265; no. 9; pp. 4884 - 4892
Main Authors:	MOSHIER, J. A, GILBERT, J. D, SKUNCA, M, DOSESCU, J, ALMODOVAR, K. M, LUK, G. D
Format:	Journal Article
Language:	English
Published:	Bethesda, MD American Society for Biochemistry and Molecular Biology 25-03-1990
Subjects:	Animals Base Sequence Biological and medical sciences Cell Line Colon - enzymology Cosmids DNA - genetics DNA - isolation & purification Fundamental and applied biological sciences. Psychology Gene Expression Gene Library Genes Genes. Genome Humans Intestinal Mucosa - enzymology L Cells - enzymology Mice Molecular and cellular biology Molecular genetics Molecular Sequence Data Nucleic Acid Hybridization Ornithine Decarboxylase - genetics Promoter Regions, Genetic Rats Restriction Mapping Sequence Homology, Nucleic Acid Transfection Human In vitro transcription Molecular hybridization Nucleotide sequence Enzyme Leukocyte Transcription promoter Gene organization Ornithine decarboxylase Binding site Gene expression Northern blotting Transfection Enzymatic activity Isolation Molecular cloning Transcription factor Southern blotting Comparative study
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Summary:	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.
Bibliography:	ObjectType-Article-1 SourceType-Scholarly Journals-1 ObjectType-Feature-2 content type line 23
ISSN:	0021-9258 1083-351X
DOI:	10.1016/S0021-9258(19)34057-8