The RS447 human megasatellite tandem repetitive sequence encodes a novel deubiquitinating enzyme with a functional promoter.
We have recently identified a tandem repetitive DNA sequence that we designated the RS447 megasatellite. In this study, we describe a functional novel deubiquitinating enzyme (USP17, 60 kDa) gene that is intronless and encoded by the RS447 repeating unit. Northern blot analysis in conjunction with 5' and 3' rapid amplification of cDNA ends confirmed the presence of poly(A)(+) containing RS447 RNA in normal cells. We also identified a functional promoter sequence as well as an open reading frame within every RS447 repeat. When USP17 was expressed in Escherichia coli, it exhibited deubiquitinating activity in vivo. An antibody against USP17 detected USP17 protein in human cells. Our results indicate that the RS447 repeating unit on this megasatellite repeat codes for and actively expresses a functional deubiquitinating enzyme. Although it is expressed ubiquitously in human tissues, USP17 exhibited a unique expression pattern in that its complementary strand is transcribed as an antisense transcript that may modulate the level of USP17 expression in the human brain.
Saitoh
Y
Y
Department of Molecular Neuroscience, The Institute of Medical Sciences, Isehara, Japan.
Miyamoto
N
N
Okada
T
T
Gondo
Y
Y
Showguchi-Miyata
J
J
Hadano
S
S
Ikeda
J E
JE
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D015152:Blotting, Northern; D015139:Blotting, Southern; D002478:Cells, Cultured; D004276:DNA, Satellite / Q000235:genetics*; D010450:Endopeptidases / Q000235:genetics* / Q000378:metabolism; D004926:Escherichia coli / Q000201:enzymology / Q000235:genetics / Q000378:metabolism; D015870:Gene Expression; D049452:Green Fluorescent Proteins; D006801:Humans; D008164:Luminescent Proteins / Q000096:biosynthesis; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction; D011401:Promoter Regions, Genetic / Q000235:genetics*; D012313:RNA / Q000378:metabolism; D011817:Rabbits; D017386:Sequence Homology, Amino Acid; D020080:Tandem Repeat Sequences; D014162:Transfection; D064570:Ubiquitin-Specific Proteases
D004276:DNA, Satellite; D008164:Luminescent Proteins; D049452:Green Fluorescent Proteins; D012313:RNA; D010450:Endopeptidases; D064570:Ubiquitin-Specific Proteases