Literature summary for 4.2.99.18 extracted from

Agger, S.A.; Lopez-Gallego, F.; Hoye, T.R.; Schmidt-Dannert, C.
Identification of sesquiterpene synthases from Nostoc punctiforme PCC 73102 and Nostoc sp. strain PCC 7120 (2008), J. Bacteriol., 190, 6084-6096.

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Cloned(Commentary)

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

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

Protein Variants	Comment	Organism
K364R	ntg1, localized to both nuclei and mitochondria	Saccharomyces cerevisiae

Localization

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 [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/ Products (Substrates)

Natural Substrates	Organism	Comment (Nat. Sub.)	Natural Products	Comment (Nat. Pro.)	Rev.
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

Organism	UniProt	Comment	Textmining
Saccharomyces cerevisiae	-	-	-

Posttranslational Modification

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 (Commentary)

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

Source Tissue	Comment	Organism	Textmining
cell culture	-	Saccharomyces cerevisiae	-

Substrates and Products (Substrate)

Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.
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

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