Structure of the human cellular retinoic acid-binding protein II gene. Early transcriptional regulation by retinoic acid.
The gene for human cellular retinoic acid-binding protein II (CRABP-II) has been cloned. It was isolated from a human placenta genomic library and is contained within one bacteriophage clone. The gene spans 6 kilobases and consists of 4 exons. One major transcription initiation site was mapped to an A residue 137 nucleotides upstream of the ATG initiation codon. The upstream region contains a TATA box and potential AP2, Sp1, and Krox-24 binding sites, as well as a direct repeat with homology to a retinoic acid-responsive element. The CRABP-II mRNA was rapidly induced within 2-6 h in cultured human skin fibroblasts by retinoic acid, reaching a plateau after 6 h of treatment. The rapid increase of CRABP-II message was mainly due to an increased rate of transcription as determined by nuclear run-on experiments. Increased transcription could be detected as early as 1 h after addition of retinoic acid, peaked at 2 h, and returned to basal levels within 6 h. On-going protein synthesis was required for this transient increase of transcription, since the induction was blocked by cycloheximide. These data suggest that the human CRABP-II gene is transcriptionally regulated by a newly synthesized regulatory protein.
Aström
A
A
Department of Dermatology, University of Michigan, Ann Arbor 48109.
Pettersson
U
U
Voorhees
J J
JJ
D000595:Amino Acid Sequence; D001483:Base Sequence; D001665:Binding Sites; D015152:Blotting, Northern; D002352:Carrier Proteins / Q000235:genetics*; D002467:Cell Nucleus / Q000187:drug effects / Q000378:metabolism; D002478:Cells, Cultured; D003062:Codon; D003513:Cycloheximide / Q000494:pharmacology; D004247:DNA / Q000235:genetics / Q000378:metabolism; D003609:Dactinomycin / Q000494:pharmacology; D005260:Female; D005347:Fibroblasts / Q000187:drug effects / Q000378:metabolism; D005786:Gene Expression Regulation / Q000187:drug effects*; D015698:Genomic Library; D006801:Humans; D008969:Molecular Sequence Data; D010920:Placenta / Q000502:physiology; D011247:Pregnancy; D012333:RNA, Messenger / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D018168:Receptors, Retinoic Acid; D015183:Restriction Mapping; D012867:Skin / Q000378:metabolism*; D016385:TATA Box; D014157:Transcription Factors / Q000378:metabolism; D014158:Transcription, Genetic / Q000187:drug effects*; D014212:Tretinoin / Q000494:pharmacology*
D002352:Carrier Proteins; D003062:Codon; D012333:RNA, Messenger; D018168:Receptors, Retinoic Acid; D014157:Transcription Factors; D003609:Dactinomycin; D014212:Tretinoin; D004247:DNA; D003513:Cycloheximide