Spermidine/spermine N1-acetyltransferase overexpression in kidney epithelial cells disrupts polyamine homeostasis, leads to DNA damage, and causes G2 arrest.
Expression of spermidine/spermine N(1)-acetyltransferase (SSAT) increases in kidneys subjected to ischemia-reperfusion injury (IRI). Increased expression of SSAT in vitro leads to alterations in cellular polyamine content, depletion of cofactors and precursors of polyamine synthesis, and reduced cell proliferation. In our model system, a >28-fold increase in SSAT levels in HEK-293 cells leads to depletion of polyamines and elevation in the enzymatic activities of ornithine decarboxylase and S-adenosylmethionine decarboxylase, suggestive of a compensatory reaction to increased polyamine catabolism. Increased expression of SSAT also led to DNA damage and G(2) arrest. The increased DNA damage was primarily due to the depletion of polyamines. Other factors such as increased production of H(2)O(2) due to polyamine oxidase activity may play a secondary role in the induction of DNA lesions. In response to DNA damage the ATM/ATR --> Chk1/2 DNA repair and cell cycle checkpoint pathways were activated, mediating the G(2) arrest in SSAT-expressing cells. In addition, the activation of ERK1 and ERK2, which play integral roles in the G(2)/M transition, is impaired in cells expressing SSAT. These results indicate that the disruption of polyamine homeostasis due to enhanced SSAT activity leads to DNA damage and reduced cell proliferation via activation of DNA repair and cell cycle checkpoint and disruption of Raf --> MEK --> ERK pathways. We propose that in kidneys subjected to IRI, one mechanism through which increased expression of SSAT may cause cellular injury and organ damage is through induction of DNA damage and the disruption of cell cycle.
Zahedi
Kamyar
K
Division of Nephrology and Hypertension, Children's Hospital Medical Center, University of Cincinnati College of Medicine, Cincinnati, OH 45267, USA.
Bissler
John J
JJ
Wang
Zhaohui
Z
Josyula
Anuradha
A
Lu
Lu
L
Diegelman
Paula
P
Kisiel
Nick
N
Porter
Carl W
CW
Soleimani
Manoocher
M
D000123:Acetyltransferases / Q000378:metabolism*; D002460:Cell Line; D004249:DNA Damage / Q000502:physiology*; D004847:Epithelial Cells / Q000166:cytology / Q000502:physiology*; D016195:G2 Phase / Q000502:physiology*; D006706:Homeostasis / Q000502:physiology; D006801:Humans; D007668:Kidney / Q000166:cytology / Q000502:physiology*; D011073:Polyamines / Q000378:metabolism*; D013095:Spermidine / Q000378:metabolism*; D015854:Up-Regulation / Q000502:physiology; D000099228:Diamine N-Acetyltransferase
D000123:Acetyltransferases; D011073:Polyamines; D013095:Spermidine; D000099228:Diamine N-Acetyltransferase