A mouse amidase specific for N-terminal asparagine - The gene, the enzyme, and their function in the N-end rule pathway

被引:72
作者
Grigoryev, S
Stewart, AE
Kwon, YT
Arfin, SM
Bradshaw, RA
Jenkins, NA
Copeland, NG
Varshavsky, A
机构
[1] CALTECH, DIV BIOL, PASADENA, CA 91125 USA
[2] UNIV CALIF IRVINE, COLL MED, DEPT BIOL CHEM, IRVINE, CA 92717 USA
[3] NCI, FREDERICK CANC RES & DEV CTR, ABL BASIC RES PROGRAM, FREDERICK, MD 21702 USA
关键词
D O I
10.1074/jbc.271.45.28521
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
The N-end rule relates the in vivo half-life of a protein to the identity of its N-terminal residue. In both fungi and mammals, the tertiary destabilizing N-terminal residues asparagine and glutamine function through their conversion, by enzymatic deamidation, into the secondary destabilizing residues aspartate and glutamate, whose destabilizing activity requires their enzymatic conjugation to arginine, one of the primary destabilizing residues. We report the isolation and analysis of a mouse cDNA and the corresponding gene (termed Ntan1) that encode a 310-residue amidohydrolase (termed Nt(N)-amidase) specific for N-terminal asparagine, The similar to 17-kilobase pair Ntan1 gene is located in the proximal region of mouse chromosome 16 and contains 10 exons ranging from 54 to 177 base pairs in length. The similar to 1.4 kilobase pair Ntan1 mRNA is expressed in all of the tested mouse tissues and cell lines and is down-regulated upon the conversion of myoblasts into myotubes. The Ntan1 promoter is located similar to 500 base pairs upstream of the Ntan1 start codon. The deduced amino acid sequence of mouse Nt(N)-amidase is 88% identical to the sequence of its porcine counterpart, but bears no significant similarity to the sequence of the NTA1-encoded N-terminal amidohydrolase of the yeast Saccharomyces cerevisiae, which can deamidate either N-terminal asparagine or glutamine. The expression of mouse Nt(N)-amidase in S. cerevisiae nta1 Delta was used to verify that Nt(N)-amidase retains its asparagine selectivity in. vivo and can implement the asparagine-specific subset of the N-end rule. Further dissection of mouse Ntan1, including its null phenotype analysis, should illuminate the functions of the N-end rule, most of which are still unknown.
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页码:28521 / 28532
页数:12
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