Yeast apurinic/apyrimidinic endonuclease Apn1 protects mammalian neuronal cell line from oxidative stress.
Reactive oxygen species (ROS) have been implicated as one of the agents responsible for many neurodegenerative diseases. A critical target for ROS is DNA. Most oxidative stress-induced DNA damage in the nucleus and mitochondria is removed by the base excision repair pathway. Apn1 is a yeast enzyme in this pathway which possesses a wider substrate specificity and greater enzyme activity than its mammalian counterpart for removing DNA damage, making it a good therapeutic candidate. For this study we targeted Apn1 to mitochondria in a neuronal cell line derived from the substantia nigra by using a mitochondrial targeting signal (MTS) in an effort to hasten the removal of DNA damage and thereby protect these cells. We found that following oxidative stress, mitochondrial DNA (mtDNA) was repaired more efficiently in cells containing Apn1 with the MTS than controls. There was no difference in nuclear repair. However, cells that expressed Apn1 without the MTS showed enhanced repair of both nuclear and mtDNA. Both Apn1-expressing cells were more resistant to cell death following oxidative stress compared with controls. Therefore, these results reveal that the expression of Apn1 in neurons may be of potential therapeutic benefit for treating patients with specific neurodegenerative diseases.
Ho
Renee
R
Department of Cell Biology and Neuroscience, College of Medicine, University of South Alabama, Mobile, Alabama 36688, USA.
Rachek
Lyudmila I
LI
Xu
Yi
Y
Kelley
Mark R
MR
LeDoux
Susan P
SP
Wilson
Glenn L
GL
D000818:Animals; D015139:Blotting, Southern; D015153:Blotting, Western; D002460:Cell Line; D002470:Cell Survival / Q000187:drug effects / Q000235:genetics / Q000502:physiology; D003114:Colony-Forming Units Assay; D020552:Comet Assay; D004260:DNA Repair / Q000187:drug effects; D045643:DNA Repair Enzymes; D004272:DNA, Mitochondrial / Q000235:genetics / Q000378:metabolism; D004706:Endodeoxyribonucleases / Q000235:genetics / Q000502:physiology*; D009474:Neurons / Q000187:drug effects*; D009841:Oligonucleotides / Q000378:metabolism; D018384:Oxidative Stress / Q000187:drug effects*; D010957:Plasmids / Q000235:genetics; D051381:Rats; D011916:Rats, Inbred F344; D029701:Saccharomyces cerevisiae Proteins / Q000235:genetics / Q000502:physiology*; D013347:Subcellular Fractions / Q000187:drug effects / Q000378:metabolism; D013379:Substrate Specificity; D014162:Transfection; D014446:Tyrosine 3-Monooxygenase / Q000096:biosynthesis
D004272:DNA, Mitochondrial; D009841:Oligonucleotides; D029701:Saccharomyces cerevisiae Proteins; D014446:Tyrosine 3-Monooxygenase; D004706:Endodeoxyribonucleases; C083274:Apn1 protein, S cerevisiae; D045643:DNA Repair Enzymes