Identification of a novel isopeptidase with dual specificity for ubiquitin- and NEDD8-conjugated proteins.
Covalent conjugation of proteins by ubiquitin or ubiquitin-like molecules is an important form of post-translational modification and plays a critical role in many cellular processes. Similar to the concept of phosphorylation and dephosphorylation, these conjugates are regulated by a large number of deconjugating enzymes. Here, we report the cloning of a 2,141-base pair DNA fragment from human placenta cDNA library by a strategy that involves expressed sequence tag data base searching, polymerase chain reaction, and rapid amplification of cDNA ends. Nucleotide sequence analysis revealed that the cloned cDNA contains an open reading frame of 1,143 base pairs encoding a novel protease, USP21, which is composed of 381 residues with a calculated molecular mass of 43 kDa. The human USP21 gene is located on chromosome 1q21 and encodes a member of the ubiquitin-specific protease family with highly conserved Cys and His domains. The activity and specificity of USP21 were determined by using a COS cell expression system in vivo. We showed that USP21 is capable of removing ubiquitin from ubiquitinated proteins as expected. Furthermore, USP21 is capable of removing NEDD8 from NEDD8 conjugates but has no effect on Sentrin-1 conjugates. As expected from its biochemical activity, overexpression of USP21 has a profound growth inhibitory effect on U2OS cells. Thus, USP21 is the first ubiquitin-specific protease shown to have dual specificity for both ubiquitin and NEDD8 and may play an important role in the regulation of cell growth.
Gong
L
L
Research Center for Cardiovascular Diseases, Institute of Molecular Medicine for the Prevention of Human Diseases, and Division of Molecular Medicine, Department of Internal Medicine, University of Texas Health Science Center, Houston, Texas 77030, USA.
Kamitani
T
T
Millas
S
S
Yeh
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ET
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D019556:COS Cells; D019759:Carbon-Nitrogen Lyases / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D002874:Chromosome Mapping; D003001:Cloning, Molecular; D020224:Expressed Sequence Tags; D006801:Humans; D008969:Molecular Sequence Data; D000075687:NEDD8 Protein; D017386:Sequence Homology, Amino Acid; D013379:Substrate Specificity; D043222:Ubiquitin Thiolesterase; D014452:Ubiquitins / Q000378:metabolism*
D000075687:NEDD8 Protein; C082860:NEDD8 protein, human; C408875:USP21 protein, human; D014452:Ubiquitins; D043222:Ubiquitin Thiolesterase; D019759:Carbon-Nitrogen Lyases