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

Sergei Grigoryeve, Albert E. Stewart, Yongtae Kwon, Stuart M. Arfin, Ralph A. Bradshaw, Nancy A. Jenkins, Neal G. Copeland, Alexander Varshavsky

Research output: Contribution to journalArticle

62 Citations (Scopus)

Abstract

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 ~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 ~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 -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 nta1Δ 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.

Original languageEnglish
Pages (from-to)28521-28532
Number of pages12
JournalJournal of Biological Chemistry
Volume271
Issue number45
DOIs
StatePublished - 19 Nov 1996

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amidase
Asparagine
Genes
Amidohydrolases
Yeast
Enzymes
Glutamine
Base Pairing
Dissection
Mammals
Saccharomyces cerevisiae
Chromosomes
Fungi
Aspartic Acid
Arginine
Chromosomes, Human, Pair 16
Glutamic Acid
Exons
Initiator Codon
Complementary DNA

Cite this

Grigoryeve, Sergei ; Stewart, Albert E. ; Kwon, Yongtae ; Arfin, Stuart M. ; Bradshaw, Ralph A. ; Jenkins, Nancy A. ; Copeland, Neal G. ; Varshavsky, Alexander. / A mouse amidase specific for N-terminal asparagine : The gene, the enzyme, and their function in the N-end rule pathway. In: Journal of Biological Chemistry. 1996 ; Vol. 271, No. 45. pp. 28521-28532.
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abstract = "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 ~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 ~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 -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 nta1Δ 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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Grigoryeve, S, Stewart, AE, Kwon, Y, Arfin, SM, Bradshaw, RA, Jenkins, NA, Copeland, NG & Varshavsky, A 1996, 'A mouse amidase specific for N-terminal asparagine: The gene, the enzyme, and their function in the N-end rule pathway', Journal of Biological Chemistry, vol. 271, no. 45, pp. 28521-28532. https://doi.org/10.1074/jbc.271.45.28521

A mouse amidase specific for N-terminal asparagine : The gene, the enzyme, and their function in the N-end rule pathway. / Grigoryeve, Sergei; Stewart, Albert E.; Kwon, Yongtae; Arfin, Stuart M.; Bradshaw, Ralph A.; Jenkins, Nancy A.; Copeland, Neal G.; Varshavsky, Alexander.

In: Journal of Biological Chemistry, Vol. 271, No. 45, 19.11.1996, p. 28521-28532.

Research output: Contribution to journalArticle

TY - JOUR

T1 - A mouse amidase specific for N-terminal asparagine

T2 - The gene, the enzyme, and their function in the N-end rule pathway

AU - Grigoryeve, Sergei

AU - Stewart, Albert E.

AU - Kwon, Yongtae

AU - Arfin, Stuart M.

AU - Bradshaw, Ralph A.

AU - Jenkins, Nancy A.

AU - Copeland, Neal G.

AU - Varshavsky, Alexander

PY - 1996/11/19

Y1 - 1996/11/19

N2 - 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 ~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 ~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 -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 nta1Δ 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.

AB - 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 ~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 ~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 -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 nta1Δ 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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U2 - 10.1074/jbc.271.45.28521

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AN - SCOPUS:0029905594

VL - 271

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JO - Journal of Biological Chemistry

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