Journal article


Intracellular glycation of nuclear DNA, mitochondrial DNA, and cytosolic proteins during senescence-like growth arrest


Publication Details
Author(s): Breyer V, Becker CM, Pischetsrieder M
Publisher: Mary Ann Liebert
Publication year: 2011
Volume: 30
Journal issue: 9
Pages range: 681-689
ISSN: 1044-5498

Abstract

To investigate the accumulation of intracellular advanced glycation end products (AGEs), a method was established for the simultaneous analysis of glycation products of cytosolic proteins, nuclear DNA, and mitochondrial DNA (mtDNA). Nuclear DNA, mtDNA, and cytosolic proteins were simultaneously isolated from one cell lysate by differential centrifugation and combined mechanical and chemical cell disruption methods. The major DNA-AGE N 2-carboxyethyl- 2′-deoxyguanosine (CEdG) was quantified in nuclear DNA and mtDNA by ELISA, whereas the protein-AGEs N-(carboxymethyl)lysine (CML) and N ε-(carboxyethyl)lysine (CEL) were determined by western blot. The method was used to analyze NIH3T3 fibroblasts. In untreated cells, CEdG levels of mtDNA (14.84 ± 3.07 pg CEdG/μg mtDNA) were significantly higher compared with nuclear DNA (4.40 ± 0.64 pg CEdG/μg DNA; p < 0.001). Then, fibroblasts were analyzed after 7 days of senescence-like growth arrest. In senescent fibroblasts, the CEdG content of nuclear DNA significantly increased by 25%. However, the CEdG level of mtDNA significantly decreased to 52%; in parallel, an increase in mitochondrial mass and mtDNA was observed. Senescence did not lead to general accumulation of protein-AGEs, but two protein bands at 32 and 34 kDa showed a significant increase in the CML/CEL modification rate (208%, p < 0.001; 196%, p = 0.0016) in senescent fibroblasts compared with control cells. © Copyright 2011, Mary Ann Liebert, Inc.



How to cite
APA: Breyer, V., Becker, C.-M., & Pischetsrieder, M. (2011). Intracellular glycation of nuclear DNA, mitochondrial DNA, and cytosolic proteins during senescence-like growth arrest. DNA and Cell Biology, 30(9), 681-689. https://dx.doi.org/10.1089/dna.2011.1236

MLA: Breyer, Viola, Cord-Michael Becker, and Monika Pischetsrieder. "Intracellular glycation of nuclear DNA, mitochondrial DNA, and cytosolic proteins during senescence-like growth arrest." DNA and Cell Biology 30.9 (2011): 681-689.

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