Human glyceraldehyde-3-phosphate dehydrogenase plays a direct role in reactivating oxidized forms of the DNA repair enzyme APE1.
Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) has diverse biological functions including its nuclear translocation in response to oxidative stress. We show that GAPDH physically associates with APE1, an essential enzyme involved in the repair of abasic sites in damaged DNA, as well as in the redox regulation of several transcription factors. This interaction allows GAPDH to convert the oxidized species of APE1 to the reduced form, thereby reactivating its endonuclease activity to cleave abasic sites. The GAPDH variants C152G and C156G retain the ability to interact with but are unable to reactivate APE1, implicating these cysteines in catalyzing the reduction of APE1. Interestingly, GAPDH-small interfering RNA knockdown sensitized the cells to methyl methane sulfonate and bleomycin, which generate lesions that are repaired by APE1, but showed normal sensitivity to 254-nm UV. Moreover, the GAPDH knockdown cells exhibited an increased level of spontaneous abasic sites in the genomic DNA as a result of diminished APE1 endonuclease activity. Thus, the nuclear translocation of GAPDH during oxidative stress constitutes a protective mechanism to safeguard the genome by preventing structural inactivation of APE1.
Azam
Sonish
S
University of Montreal, Maisonneuve-Rosemont Hospital, Research Center, Montreal, Quebec H1T 2M4, Canada.
Jouvet
Nathalie
N
Jilani
Arshad
A
Vongsamphanh
Ratsavarinh
R
Yang
Xiaoming
X
Yang
Stephen
S
Ramotar
Dindial
D
D021581:Active Transport, Cell Nucleus / Q000187:drug effects / Q000528:radiation effects; D000903:Antibiotics, Antineoplastic / Q000494:pharmacology; D001761:Bleomycin / Q000494:pharmacology; D045744:Cell Line, Tumor; D002467:Cell Nucleus / Q000201:enzymology*; D003545:Cysteine / Q000378:metabolism; D004260:DNA Repair / Q000187:drug effects / Q000502:physiology* / Q000528:radiation effects; D043603:DNA-(Apurinic or Apyrimidinic Site) Lyase / Q000378:metabolism*; D004789:Enzyme Activation / Q000187:drug effects / Q000502:physiology / Q000528:radiation effects; D015894:Genome, Human / Q000502:physiology*; D005987:Glyceraldehyde-3-Phosphate Dehydrogenases / Q000037:antagonists & inhibitors / Q000378:metabolism*; D006801:Humans; D008741:Methyl Methanesulfonate / Q000494:pharmacology; D009153:Mutagens / Q000494:pharmacology; D010084:Oxidation-Reduction / Q000187:drug effects / Q000528:radiation effects; D018384:Oxidative Stress / Q000187:drug effects / Q000502:physiology* / Q000528:radiation effects; D034741:RNA, Small Interfering / Q000494:pharmacology; D014466:Ultraviolet Rays
D000903:Antibiotics, Antineoplastic; D009153:Mutagens; D034741:RNA, Small Interfering; D001761:Bleomycin; D008741:Methyl Methanesulfonate; D005987:Glyceraldehyde-3-Phosphate Dehydrogenases; C085108:APEX1 protein, human; D043603:DNA-(Apurinic or Apyrimidinic Site) Lyase; D003545:Cysteine
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