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Literature summary for 2.7.1.160 extracted from

  • Harding, H.P.; Lackey, J.G.; Hsu, H.C.; Zhang, Y.; Deng, J.; Xu, R.M.; Damha, M.J.; Ron, D.
    An intact unfolded protein response in Trpt1 knockout mice reveals phylogenic divergence in pathways for RNA ligation (2008), RNA, 14, 225-232.
    View publication on PubMedView publication on EuropePMC

Application

Application Comment Organism
additional information Inhibitors of Tpt1p might prove useful as selective antifungal agents. Mus musculus

Cloned(Commentary)

Cloned (Comment) Organism
The murine Trpt1 gene is targeted in E14 ES cells with a positive/negative selection vector. Trpt1 gene is disrupted by homologous recombination in ES cells, deleting exons 3, 4, and part of exon 5 that encode amino acids 48-173. Clones from two individually selected heterozygous ES cell lines are injected into blastocysts and the resultant founder chimeras bred to obtain two lines of Trpt1+/- and Trpt1-/-mice that are both viable and healthy and are normally fertile. Cell culture: Trpt1+/+ and Trpt1-/-cell lines are obtained by serial passage of SV40-T-antigen transfected mouse embryonic fibroblasts. Mus musculus

Metals/Ions

Metals/Ions Comment Organism Structure
Mg2+ 5 mM in the activity assay Mus musculus

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
2'-phospho-(ligated tRNA) + NAD+ Mus musculus
-
mature tRNA + ADP-ribose 1'',2''-phosphate + nicotinamide + H+
-
?
additional information Mus musculus Comparison of incorporation of 3H-tyrosine and 35S-methionine/cysteine into total proteins confirms that wild-type and Trpt1-/-cells have similar incorporation rates of either amino acid. ?
-
?

Organism

Organism UniProt Comment Textmining
Mus musculus
-
inactivation of the enzyme is done in knockout mice
-

Source Tissue

Source Tissue Comment Organism Textmining
blastocyst Clones from two individually selected heterozygous ES cell lines are injected into blastocysts and the resultant founder chimeras bred to obtain two lines of knockout Trpt1+/- and knockout Trpt1-/-mice. Mus musculus
-
heart
-
Mus musculus
-
MEF cell Trpt1+/+ and Trpt1-/-cell lines are obtained by serial passage of SV40-T-antigen transfected mouse embryonic fibroblasts Mus musculus
-

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
additional information
-
Despite lack of detectable in vitro RNA 2'-phosphotransferase activity in lysates of cells derived from the Trpt1 knockout mice, there is no evidence for a defect in the unfolded protein response or in tRNA function. Mus musculus
additional information
-
Immunoprecipitation of Trpt1 from heart lysates, followed by immunoblot with an antiserum raised to bacterially expressed murine Trpt1 reveals that Trpt1-/-mice lack 2'-phosphotransferase activity yet retain the ability to translate spliced transcription factor XBP-1 mRNA and synthesize tyrosine-containing proteins at wild-type levels. Heterozygotes express 50% of wild-type levels. Extracts of Trpt1+/+cells have measurable NAD+-dependent 2'-phosphotransferase activity. Mus musculus

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
2'-phospho-(ligated tRNA) + NAD+
-
Mus musculus mature tRNA + ADP-ribose 1'',2''-phosphate + nicotinamide + H+
-
?
additional information Comparison of incorporation of 3H-tyrosine and 35S-methionine/cysteine into total proteins confirms that wild-type and Trpt1-/-cells have similar incorporation rates of either amino acid. Mus musculus ?
-
?

Synonyms

Synonyms Comment Organism
2'-phosphotransferase
-
Mus musculus
Tpt1p Mammalian genomes encode a single known functional homolog of yeast Tpt1p. Mus musculus
Trpt1
-
Mus musculus

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
30
-
assay at Mus musculus

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
7.5
-
assay at Mus musculus

Cofactor

Cofactor Comment Organism Structure
NAD+
-
Mus musculus