Linked Life Data

9653654

Source:http://linkedlifedata.com/resource/pubmed/id/9653654

Statements in which the resource exists as a subject.
PredicateObject
rdf:type
lifeskim:mentions
pubmed:issue
2
pubmed:dateCreated
1998-10-1
pubmed:databankReference
pubmed:abstractText
The full-length cDNA and genomic sequences encoding murine acid ceramidase (AC; E.C. 3.5.1.23) have been isolated and characterized. The 2176-bp cDNA was approximately 80% identical to the human cDNA (Koch et al., 1996) and predicted a 394-amino-acid polypeptide that was approximately 90% identical to the human protein. A fluorescence-based assay system was developed to determine AC enzymatic activity, and transfection of COS-1 cells with the full-length mouse cDNA led to increased AC activity, demonstrating its functionality. The murine AC gene, which spanned approximately 38 kb, consisted of 14 exons separated by 13 introns. The exons ranged in size from 46 to 1038 bp and were flanked by exon/intron junctions that adhered closely to known donor and acceptor splice site consensus sequences. Exon 1 encoded the putative translation start site and the signal peptide region, while exon 14 encoded the carboxy end of the AC polypeptide and all of the 3' untranslated region. Sequence analysis of a 497-bp region upstream from the first in-frame ATG revealed several features of a housekeeping promoter, as well as several tissue-specific and/or hormone-inducible regulatory sites. Insertion of this sequence into a chloramphenicol acyltransferase (CAT) expression vector led an approximately fivefold increase in CAT activity after transfection into NIH3T3 cells. Northern blot analysis and enzymatic assays also were carried out on various murine tissues to examine AC expression. Of the tissues studied, the highest AC activity and mRNA levels were found in the kidney, followed by the brain; almost no AC activity or mRNA was found in the testis or skeletal muscle. These latter studies provided clear evidence that despite the housekeeping function of AC, its expression was tissue-specific.
pubmed:grant
pubmed:language
eng
pubmed:journal
pubmed:citationSubset
IM
pubmed:chemical
pubmed:status
MEDLINE
pubmed:month
Jun
pubmed:issn
0888-7543
pubmed:author
pubmed:issnType
Print
pubmed:day
1
pubmed:volume
50
pubmed:owner
NLM
pubmed:authorsComplete
Y
pubmed:pagination
267-74
pubmed:dateRevised
2008-11-21
pubmed:meshHeading
pubmed-meshheading:9653654-Acid Ceramidase, pubmed-meshheading:9653654-Amidohydrolases, pubmed-meshheading:9653654-Amino Acid Sequence, pubmed-meshheading:9653654-Animals, pubmed-meshheading:9653654-Base Sequence, pubmed-meshheading:9653654-Blotting, Northern, pubmed-meshheading:9653654-Brain, pubmed-meshheading:9653654-Cell Line, pubmed-meshheading:9653654-Ceramidases, pubmed-meshheading:9653654-Cloning, Molecular, pubmed-meshheading:9653654-Consensus Sequence, pubmed-meshheading:9653654-DNA, Complementary, pubmed-meshheading:9653654-Exons, pubmed-meshheading:9653654-Gene Expression, pubmed-meshheading:9653654-Humans, pubmed-meshheading:9653654-Introns, pubmed-meshheading:9653654-Kidney, pubmed-meshheading:9653654-Mice, pubmed-meshheading:9653654-Molecular Sequence Data, pubmed-meshheading:9653654-Mutagenesis, Site-Directed, pubmed-meshheading:9653654-Promoter Regions, Genetic, pubmed-meshheading:9653654-RNA, Messenger, pubmed-meshheading:9653654-Sequence Analysis, DNA, pubmed-meshheading:9653654-Transfection
pubmed:year
1998
pubmed:articleTitle
Cloning and characterization of the full-length cDNA and genomic sequences encoding murine acid ceramidase.
pubmed:affiliation
Department of Human Genetics, Mount Sinai School of Medicine, New York, New York 10029, USA.
pubmed:publicationType
Journal Article, Research Support, U.S. Gov't, P.H.S., Research Support, U.S. Gov't, Non-P.H.S., Research Support, Non-U.S. Gov't