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rdf:type |
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lifeskim:mentions |
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pubmed:issue |
12
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pubmed:dateCreated |
2002-3-18
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pubmed:abstractText |
All-trans-retinoic acid (ATRA) induces myeloid differentiation of a human promyelocytic leukemia cell line, NB4, but does not affect its subclone NB4/RA harboring a point-mutated ligand-binding domain (AF2) in retinoic acid receptor alpha (RARalpha) gene. We found that ATRA induced the 4-fold elevation of acid sphingomyelinase (ASMase) activity 24 h after treatment in NB4 cells, but not in NB4/RA cells. ATRA did not affect neutral sphingomyelinase activity in either NB4 or NB4/RA. Upon treatment with ATRA, ceramide, the product of an ASMase reaction, accumulated in NB4 cells. Northern blot analysis showed a marked elevation of the ASMase mRNA 8 h after ATRA treatment, reaching a plateau at 24 h. Regulation of ASMase gene expression was studied by a promoter analysis using luciferase reporter assay. The 5'-upstream flanking region of human ASMase gene (-519/+300) conjugated with the luciferase gene was introduced into COS-7 cells. Luciferase activity in transformed cells markedly increased in response to ATRA stimulation when the wild type RARalpha or the PML/RARalpha hybrid protein was co-expressed. Deletion experiments revealed that a short sequence at the 5'-end (-519/-485) was indispensable for the ATRA response. Within this short region, two retinoic acid-responsive element-like motifs (TGCCCG and TCTCCT) and one AP2-like motif (CCCTTCCC) were identified. Deletion and base-substitution experiments showed that all three motifs are required for the full expression induced by ATRA. Electrophoresis mobility shift assays with the nuclear extract of ATRA-treated NB4 cells showed that proteins were bound specifically to the probe being mediated by all three motifs in the promoter sequence.
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pubmed:language |
eng
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pubmed:journal |
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pubmed:citationSubset |
IM
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pubmed:chemical |
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pubmed:status |
MEDLINE
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pubmed:month |
Mar
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pubmed:issn |
0021-9258
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pubmed:author |
pubmed-author:AsanoHaruhikoH,
pubmed-author:HattoriMasashiM,
pubmed-author:HottaTomomitsuT,
pubmed-author:KojimaTetsuhitoT,
pubmed-author:MurateTakashiT,
pubmed-author:SaitoHidehikoH,
pubmed-author:SuzukiMotoshiM,
pubmed-author:TakagiAkiraA,
pubmed-author:Tamiya-KoizumiKeikoK,
pubmed-author:TanizawaTomomiT,
pubmed-author:YoshidaShonenS
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pubmed:issnType |
Print
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pubmed:day |
22
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pubmed:volume |
277
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pubmed:owner |
NLM
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pubmed:authorsComplete |
Y
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pubmed:pagination |
9936-43
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pubmed:dateRevised |
2006-5-1
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pubmed:meshHeading |
pubmed-meshheading:11788605-Amino Acid Motifs,
pubmed-meshheading:11788605-Animals,
pubmed-meshheading:11788605-Base Sequence,
pubmed-meshheading:11788605-Blotting, Northern,
pubmed-meshheading:11788605-COS Cells,
pubmed-meshheading:11788605-Cell Differentiation,
pubmed-meshheading:11788605-Cell Nucleus,
pubmed-meshheading:11788605-Ceramides,
pubmed-meshheading:11788605-Dose-Response Relationship, Drug,
pubmed-meshheading:11788605-Gene Deletion,
pubmed-meshheading:11788605-Genes, Reporter,
pubmed-meshheading:11788605-Humans,
pubmed-meshheading:11788605-Leukemia, Promyelocytic, Acute,
pubmed-meshheading:11788605-Luciferases,
pubmed-meshheading:11788605-Molecular Sequence Data,
pubmed-meshheading:11788605-Mutation,
pubmed-meshheading:11788605-Plasmids,
pubmed-meshheading:11788605-Protein Binding,
pubmed-meshheading:11788605-Protein Structure, Tertiary,
pubmed-meshheading:11788605-Receptors, Retinoic Acid,
pubmed-meshheading:11788605-Reverse Transcriptase Polymerase Chain Reaction,
pubmed-meshheading:11788605-Spectrometry, Mass, Electrospray Ionization,
pubmed-meshheading:11788605-Sphingomyelin Phosphodiesterase,
pubmed-meshheading:11788605-Time Factors,
pubmed-meshheading:11788605-Transfection,
pubmed-meshheading:11788605-Tretinoin,
pubmed-meshheading:11788605-Tumor Cells, Cultured,
pubmed-meshheading:11788605-Up-Regulation,
pubmed-meshheading:11788605-beta-Galactosidase
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pubmed:year |
2002
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pubmed:articleTitle |
Up-regulation of acid sphingomyelinase during retinoic acid-induced myeloid differentiation of NB4, a human acute promyelocytic leukemia cell line.
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pubmed:affiliation |
Nagoya University School of Health Science, Nagoya 466-8550, Japan. murate@met.nagoya-u.ac.jp
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pubmed:publicationType |
Journal Article
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