Cloned (Comment) | Organism |
---|---|
The pPS904 green fluorescent protein (GFP) expression vector (2m URA3) is employed for generation of C-terminally tagged Ntg1-GFP fusion protein. The Saccharomyces cerevisiae haploid strain FY86 is utilized for all localization studies. NTG1 strain (DSC0282) is generated by precisely replacing the NTG1 open reading frame in FY86 with the kanamycin antibiotic resistance gene. Haploid yeast strain expresses integrated genomic copies of C-terminally tandem affinity purification (TAP)-tagged Ntg1. Cells expressing galactose-inducible SMT3-HA and Ntg1-GST (DSC0221) are generated by integrating the hemagglutinin tag from the vector p1375 and the GAL promoter and glutathione S-transferase (GST) tag from the vector p2245 at the C-termini of the SMT3 and NTG1 products in the haploid strain ACY737. | Saccharomyces cerevisiae |
The pPS904 green fluorescent protein (GFP) expression vector (2m URA3) is employed for generation of C-terminally tagged Ntg2-GFP fusion protein. The Saccharomyces cerevisiae haploid strain FY86 is utilized for localization studies. NTG2 strain (DSC0283) is generated by precisely replacing NTG2 open reading frame in FY86 with the kanamycin antibiotic resistance gene. Haploid yeast strain expresses integrated genomic copies of C-terminally tandem affinity purification (TAP)-tagged Ntg2. Cells expressing galactose-inducible Smt3-HA and Ntg2-GST (DSC0222) are generated by integrating the hemagglutinin tag from the vector p1375 and the GAL promoter and glutathione S-transferase (GST) tag from the vector p2245 at the C-termini of the SMT3 and NTG2 products in the haploid strain ACY737. | Saccharomyces cerevisiae |
Protein Variants | Comment | Organism |
---|---|---|
K364R | ntg1, localized to both nuclei and mitochondria | Saccharomyces cerevisiae |
Localization | Comment | Organism | GeneOntology No. | Textmining |
---|---|---|---|---|
mitochondrion | - |
Saccharomyces cerevisiae | 5739 | - |
additional information | localization of Ntg1 is dynamically regulated in response to nuclear and mitochondrial oxidative stress | Saccharomyces cerevisiae | - |
- |
nucleus | - |
Saccharomyces cerevisiae | 5634 | - |
nucleus | sumoylated Ntg1 accumulates in the nucleus following oxidative stress | Saccharomyces cerevisiae | 5634 | - |
Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
---|---|---|---|
46000 | - |
Ntg2-TAP, affirmed by Western Blot analysis | Saccharomyces cerevisiae |
55000 | - |
calculated for Ntg1-TAP, affirmed by Western Blot analysis | Saccharomyces cerevisiae |
58000 | - |
monosumoylated Ntg2, affirmed by Western Blot analysis | Saccharomyces cerevisiae |
70000 | - |
monosumoylated Ntg1-TAP, affirmed by Western Blot analysis | Saccharomyces cerevisiae |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
DNA | Saccharomyces cerevisiae | oxidative DNA damage is primarily reversed by the base excision repair pathway, initiated by N-glycosylase apurinic/apyrimidinic lyase proteins | fragments of DNA | - |
ir | |
additional information | Saccharomyces cerevisiae | ntg1 possesses N-glycosylase/AP lyase activity that allows recognition and repair of oxidative base damage (primarily of pyrimidines) as well as abasic sites | ? | - |
? | |
additional information | Saccharomyces cerevisiae | ntg2 possesses N-glycosylase/AP lyase activity that allows recognition and repair of oxidative base damage (primarily of pyrimidines) as well as abasic sites | ? | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Saccharomyces cerevisiae | - |
- |
- |
Posttranslational Modification | Comment | Organism |
---|---|---|
sumoylation | modification with the ubiquitin-like SUMO protein | Saccharomyces cerevisiae |
sumoylation | modification with the ubiquitin-like SUMO protein, is associated with targeting of Ntg1 to nuclei containing oxidative DNA damage, lysine 364 within the sequence KREL is most likely to be sumoylated among the 36 lysines present in Ntg1 | Saccharomyces cerevisiae |
Purification (Comment) | Organism |
---|---|
TAP-tagged Ntg1 is purified by a modified TAP method. To purify GST-tagged Ntg1, glutathione-Sepharose 4 Fast Flow beads are applied | Saccharomyces cerevisiae |
TAP-tagged Ntg2 is purified by a modified TAP method. To purify GST-tagged Ntg2, glutathione-Sepharose 4 Fast Flow beads are applied | Saccharomyces cerevisiae |
Source Tissue | Comment | Organism | Textmining |
---|---|---|---|
cell culture | - |
Saccharomyces cerevisiae | - |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
DNA | oxidative DNA damage is primarily reversed by the base excision repair pathway, initiated by N-glycosylase apurinic/apyrimidinic lyase proteins | Saccharomyces cerevisiae | fragments of DNA | - |
ir | |
additional information | ntg1 possesses N-glycosylase/AP lyase activity that allows recognition and repair of oxidative base damage (primarily of pyrimidines) as well as abasic sites | Saccharomyces cerevisiae | ? | - |
? | |
additional information | ntg2 possesses N-glycosylase/AP lyase activity that allows recognition and repair of oxidative base damage (primarily of pyrimidines) as well as abasic sites | Saccharomyces cerevisiae | ? | - |
? |
Synonyms | Comment | Organism |
---|---|---|
endonuclease III | - |
Saccharomyces cerevisiae |
N-glycosylase AP lyase | - |
Saccharomyces cerevisiae |
N-glycosylase apurinic/apyrimidinic lyase | - |
Saccharomyces cerevisiae |
Ntg1 | - |
Saccharomyces cerevisiae |
Ntg2 | - |
Saccharomyces cerevisiae |