Characterization of the 5'-flanking region of the human tartrate-resistant acid phosphatase (TRAP) gene.
Tartrate-resistant acid phosphatase (TRAP) is expressed at high levels in osteoclasts and may play an important role in the bone resorptive process. However, factors regulating human TRAP gene expression have not been clearly defined. Therefore, we isolated a genomic clone (CL-9) for TRAP containing a 14-kb insert. A restriction map was generated for this insert, and a 2.6-kb ApaI fragment containing the 5'-flanking region was subcloned. Sequence analysis of this fragment revealed the presence of candidate transcription factor-binding sequences for H-APF-1, SP1, GATA2, and the c-Myc proto-oncogene. PCR analysis of RNA isolated from human osteoclastomas and pagetic bone revealed a 276-bp intron at -1 bp to -276 bp relative to the ATG and a transcript originating from this intron. Rapid amplification of the 5' end of the human TRAP mRNA by PCR indicated the presence of a 93-bp untranslated region 5' from the intron. Promoter activity was detected in the DNA fragment from +1 bp to -1903 bp relative to the ATG initiation codon, which drove the transient expression of a luciferase reporter gene when transfected into HRE H9 rabbit endometrial cells. Comparison of the human TRAP 5'-flanking region with mouse TRAP and uteroferrin revealed 41% and 47% homology, respectively. This suggests that regulation of human TRAP gene expression may differ from that for the murine TRAP gene.
Reddy
S V
SV
Department of Medicine/Hematology, Audie Murphy Veterans Administration Hospital, University of Texas Health Science Center, San Antonio 78284, USA.
Kuzhandaivelu
N
N
Acosta
L G
LG
Roodman
G D
GD
D000135:Acid Phosphatase / Q000187:drug effects / Q000235:genetics*; D000818:Animals; D001483:Base Sequence; D001862:Bone Resorption / Q000201:enzymology; D002460:Cell Line; D003001:Cloning, Molecular; D016384:Consensus Sequence; D004247:DNA / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D004717:Endometrium / Q000166:cytology / Q000378:metabolism; D005260:Female; D015971:Gene Expression Regulation, Enzymologic / Q000235:genetics*; D017930:Genes, Reporter; D006801:Humans; D007527:Isoenzymes / Q000187:drug effects / Q000235:genetics*; D008156:Luciferases / Q000235:genetics; D008667:Metalloproteins / Q000235:genetics; D008969:Molecular Sequence Data; D010010:Osteoclasts / Q000201:enzymology*; D016133:Polymerase Chain Reaction; D011401:Promoter Regions, Genetic / Q000502:physiology; D000090063:Proto-Oncogene Mas; D016271:Proto-Oncogene Proteins c-myc / Q000235:genetics / Q000378:metabolism; D011817:Rabbits; D015183:Restriction Mapping; D012689:Sequence Homology, Nucleic Acid; D000071681:Tartrate-Resistant Acid Phosphatase; D014157:Transcription Factors / Q000235:genetics / Q000378:metabolism
D007527:Isoenzymes; C000715470:MAS1 protein, human; D008667:Metalloproteins; D000090063:Proto-Oncogene Mas; D016271:Proto-Oncogene Proteins c-myc; D014157:Transcription Factors; D004247:DNA; D008156:Luciferases; D000135:Acid Phosphatase; C000605511:Acp5 protein, mouse; D000071681:Tartrate-Resistant Acid Phosphatase