A critical step in the synthesis of unsaturated fatty acids is catalysed by stearoyl-CoA desaturase (Scd). To determine the regulation of human Scd, we characterized the gene and its transcripts. Screening a human keratinocyte cDNA library and analysis of 3ʹ-RACE (rapid amplification of cDNA ends) products from various tissues yielded a 5.2 kb cDNA encoding a 359 amino acid protein with a calculated molecular mass of 41.5 kDa. Analysis of 3ʹ-RACE products suggested that alternative usage of polyadenylation sites generates two transcripts of 3.9 and 5.2 kb, a result consistent with Northern analysis. Southern analysis demonstrated the existance of two SCD loci in the human genome. Chromosomal mapping localized one locus to chromosome 10, and the second locus to chromosome 17. Characterization of genomic clones isolated from chromosome-specific libraries revealed that only the locus on chromosome 10 contained introns. Sequence analysis of the intron-less locus displayed multiple nucleotide insertions and deletions, as well as in-frame stop codons. Reverse transcriptase-PCR analysis performed with primers specific to the intron-less locus failed to produce a PCR product from brain, liver and skin RNA, indicating that the locus on chromosome 17 is most likely a transcriptionally inactive, fully processed pseudogene. These results suggest strongly that there is one structural SCD gene in the human genome, and that it generates two transcripts by use of alternative polyadenyation sites. Although the primary sequence and intron-exon structure of SCD is phylogenetically conserved, divergence between rodent and human is seen in the number of SCD genes and in the generation of alternative transcripts, suggesting a species-specific component of SCD regulation and function.
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Research Article|
May 10 1999
Human stearoyl-CoA desaturase: alternative transcripts generated from a single gene by usage of tandem polyadenylation sites
Lin ZHANG;
Lin ZHANG
1Skin Biology T. R. C., Johnson and Johnson, C. P. W. W., Skillman, NJ 08558, U.S.A.
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Lan GE;
Lan GE
1Skin Biology T. R. C., Johnson and Johnson, C. P. W. W., Skillman, NJ 08558, U.S.A.
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Satish PARIMOO;
Satish PARIMOO
1Skin Biology T. R. C., Johnson and Johnson, C. P. W. W., Skillman, NJ 08558, U.S.A.
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Kurt STENN;
Kurt STENN
1Skin Biology T. R. C., Johnson and Johnson, C. P. W. W., Skillman, NJ 08558, U.S.A.
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Stephen M. PROUTY
Stephen M. PROUTY
1
1To whom correspondence should be addressed (e-mail sprouty@cpcus.jnj.com).
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Publisher: Portland Press Ltd
Received:
November 02 1998
Revision Received:
January 20 1999
Accepted:
February 23 1999
Online ISSN: 1470-8728
Print ISSN: 0264-6021
The Biochemical Society, London © 1999
1999
Biochem J (1999) 340 (1): 255–264.
Article history
Received:
November 02 1998
Revision Received:
January 20 1999
Accepted:
February 23 1999
Citation
Lin ZHANG, Lan GE, Satish PARIMOO, Kurt STENN, Stephen M. PROUTY; Human stearoyl-CoA desaturase: alternative transcripts generated from a single gene by usage of tandem polyadenylation sites. Biochem J 15 May 1999; 340 (1): 255–264. doi: https://doi.org/10.1042/bj3400255
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