Gene structure, alternate splicing, tissue distribution, cellular localization, and developmental expression pattern of mouse deubiquitinating enzyme isoforms Usp2-45 and Usp2-69.
We have identified a novel mouse gene, Usp2, encoding two ubiquitin-specific proteases (USPs) due to alternate splicing of 5' exons. Usp2-45 consists of 396 amino acids (45.2 kDa), while Usp2-69 is 619 amino acids (69.5 kDa). Usp2-69 results from the splicing of different combinations of untranslated 5' exons (1A, 1B, 1C) onto exon 1D and the 40-kDa catalytic core (exons 3-13), while Usp2-45 has exon 2 spliced onto the core. The catalytic core contains the highly conserved motifs of the UBP family of deubiquitinating enzymes. We can find no evidence for a reported 41-kDa isoform (UBP41) in any sequence databases. Usp2-69 is able to form a complex with Usp2-45 and with itself. Antibodies raised against the catalytic core recognized a 69-kDa protein, but did not detect a 45-kDa protein in mouse tissues. Using Northern blot, Western blot, and immunohistochemistry, Usp2 expression was observed in many adult and embryonic tissues including testis, heart, skeletal muscle, diaphragm, brain, kidney, liver, pancreas, lung, and skin. Both Usp2 isoforms were localized to the cytoplasm when overexpressed in COS-7 and NIH3T3 cells. The Usp2 expression pattern indicates that this protein might be involved in specific processes in different types of cells, especially those that are differentiating, and that its function is not restricted to a development of a particular organ.
Gousseva
Natalia
N
Ubiquitin Laboratory, Division of Molecular Bioscience, John Curtin School of Medical Research, Australian National University, GPO Box 334, Canberra, ACT 2601, Australia.
Baker
Rohan T
RT
D016475:3T3 Cells / Q000201:enzymology; D017398:Alternative Splicing; D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D019556:COS Cells / Q000201:enzymology; D003001:Cloning, Molecular; D010450:Endopeptidases / Q000235:genetics* / Q000378:metabolism*; D005091:Exons; D018507:Gene Expression Regulation, Developmental; D015971:Gene Expression Regulation, Enzymologic; D007438:Introns; D051379:Mice; D008969:Molecular Sequence Data; D020033:Protein Isoforms; D022821:RNA Splice Sites; D012333:RNA, Messenger / Q000378:metabolism; D014018:Tissue Distribution; D025801:Ubiquitin / Q000378:metabolism; D043222:Ubiquitin Thiolesterase; D064570:Ubiquitin-Specific Proteases
D020033:Protein Isoforms; D022821:RNA Splice Sites; D012333:RNA, Messenger; D025801:Ubiquitin; D010450:Endopeptidases; D043222:Ubiquitin Thiolesterase; D064570:Ubiquitin-Specific Proteases; C579224:Usp2 protein, mouse