Increased TFAM binding to mtDNA damage hot spots is associated with mtDNA loss in aged rat heart

Guglielmina Chimienti, Anna Picca, Giuseppe Sirago, Flavio Fracasso, Riccardo Calvani, Roberto Bernabei, Francesco Russo, Christy S Carter, Christiaan Leeuwenburgh, Vito Pesce, Emanuele Marzetti, Angela Maria Serena Lezza

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Abstract

The well-known age-related mitochondrial dysfunction deeply affects heart because of the tissue's large dependence on mitochondrial ATP provision. Our study revealed in aged rat heart a significant 25% decrease in mtDNA relative content, a significant 29% increase in the 4.8 Kb mtDNA deletion relative content, and a significant inverse correlation between such contents as well as a significant 38% decrease in TFAM protein amount. The TFAM-binding activity to specific mtDNA regions increased at those encompassing the mtDNA replication origins, D-loop and Ori-L. The same mtDNA regions were screened for different kinds of oxidative damage, namely Single Strand Breaks (SSBs), Double Strand Breaks (DSBs), abasic sites (AP sites) and oxidized bases as 7,8-dihydro-8-oxoguanine (8oxoG). A marked increase in the relative content of mtDNA strand damage (SSBs, DSBs and AP sites) was found in the D-loop and Ori-L regions in the aged animals, unveiling for the first time in vivo an age-related, non-stochastic accumulation of oxidative lesions in these two regions that appear as hot spots of mtDNA damage. The use of Formamidopyrimidine glycosylase (Fpg) demonstrated also a significant age-related accumulation of oxidized purines particularly in the D-loop and Ori-L regions. The detected increased binding of TFAM to the mtDNA damage hot spots in aged heart suggests a link between TFAM binding to mtDNA and loss of mitochondrial genome likely through hindrance of repair processes.

Original languageEnglish
Pages (from-to)447-453
Number of pages7
JournalFree Radical Biology and Medicine
Volume124
DOIs
Publication statusPublished - Aug 20 2018

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Mitochondrial DNA
Rats
Replication Origin
Purines
Mitochondrial Genome
Animals
Repair
Genes
Adenosine Triphosphate
Tissue

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Chimienti, G., Picca, A., Sirago, G., Fracasso, F., Calvani, R., Bernabei, R., ... Lezza, A. M. S. (2018). Increased TFAM binding to mtDNA damage hot spots is associated with mtDNA loss in aged rat heart. Free Radical Biology and Medicine, 124, 447-453. https://doi.org/10.1016/j.freeradbiomed.2018.06.041

Increased TFAM binding to mtDNA damage hot spots is associated with mtDNA loss in aged rat heart. / Chimienti, Guglielmina; Picca, Anna; Sirago, Giuseppe; Fracasso, Flavio; Calvani, Riccardo; Bernabei, Roberto; Russo, Francesco; Carter, Christy S; Leeuwenburgh, Christiaan; Pesce, Vito; Marzetti, Emanuele; Lezza, Angela Maria Serena.

In: Free Radical Biology and Medicine, Vol. 124, 20.08.2018, p. 447-453.

Research output: Contribution to journalArticle

Chimienti, G, Picca, A, Sirago, G, Fracasso, F, Calvani, R, Bernabei, R, Russo, F, Carter, CS, Leeuwenburgh, C, Pesce, V, Marzetti, E & Lezza, AMS 2018, 'Increased TFAM binding to mtDNA damage hot spots is associated with mtDNA loss in aged rat heart', Free Radical Biology and Medicine, vol. 124, pp. 447-453. https://doi.org/10.1016/j.freeradbiomed.2018.06.041
Chimienti, Guglielmina ; Picca, Anna ; Sirago, Giuseppe ; Fracasso, Flavio ; Calvani, Riccardo ; Bernabei, Roberto ; Russo, Francesco ; Carter, Christy S ; Leeuwenburgh, Christiaan ; Pesce, Vito ; Marzetti, Emanuele ; Lezza, Angela Maria Serena. / Increased TFAM binding to mtDNA damage hot spots is associated with mtDNA loss in aged rat heart. In: Free Radical Biology and Medicine. 2018 ; Vol. 124. pp. 447-453.
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AU - Fracasso, Flavio

AU - Calvani, Riccardo

AU - Bernabei, Roberto

AU - Russo, Francesco

AU - Carter, Christy S

AU - Leeuwenburgh, Christiaan

AU - Pesce, Vito

AU - Marzetti, Emanuele

AU - Lezza, Angela Maria Serena

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N2 - The well-known age-related mitochondrial dysfunction deeply affects heart because of the tissue's large dependence on mitochondrial ATP provision. Our study revealed in aged rat heart a significant 25% decrease in mtDNA relative content, a significant 29% increase in the 4.8 Kb mtDNA deletion relative content, and a significant inverse correlation between such contents as well as a significant 38% decrease in TFAM protein amount. The TFAM-binding activity to specific mtDNA regions increased at those encompassing the mtDNA replication origins, D-loop and Ori-L. The same mtDNA regions were screened for different kinds of oxidative damage, namely Single Strand Breaks (SSBs), Double Strand Breaks (DSBs), abasic sites (AP sites) and oxidized bases as 7,8-dihydro-8-oxoguanine (8oxoG). A marked increase in the relative content of mtDNA strand damage (SSBs, DSBs and AP sites) was found in the D-loop and Ori-L regions in the aged animals, unveiling for the first time in vivo an age-related, non-stochastic accumulation of oxidative lesions in these two regions that appear as hot spots of mtDNA damage. The use of Formamidopyrimidine glycosylase (Fpg) demonstrated also a significant age-related accumulation of oxidized purines particularly in the D-loop and Ori-L regions. The detected increased binding of TFAM to the mtDNA damage hot spots in aged heart suggests a link between TFAM binding to mtDNA and loss of mitochondrial genome likely through hindrance of repair processes.

AB - The well-known age-related mitochondrial dysfunction deeply affects heart because of the tissue's large dependence on mitochondrial ATP provision. Our study revealed in aged rat heart a significant 25% decrease in mtDNA relative content, a significant 29% increase in the 4.8 Kb mtDNA deletion relative content, and a significant inverse correlation between such contents as well as a significant 38% decrease in TFAM protein amount. The TFAM-binding activity to specific mtDNA regions increased at those encompassing the mtDNA replication origins, D-loop and Ori-L. The same mtDNA regions were screened for different kinds of oxidative damage, namely Single Strand Breaks (SSBs), Double Strand Breaks (DSBs), abasic sites (AP sites) and oxidized bases as 7,8-dihydro-8-oxoguanine (8oxoG). A marked increase in the relative content of mtDNA strand damage (SSBs, DSBs and AP sites) was found in the D-loop and Ori-L regions in the aged animals, unveiling for the first time in vivo an age-related, non-stochastic accumulation of oxidative lesions in these two regions that appear as hot spots of mtDNA damage. The use of Formamidopyrimidine glycosylase (Fpg) demonstrated also a significant age-related accumulation of oxidized purines particularly in the D-loop and Ori-L regions. The detected increased binding of TFAM to the mtDNA damage hot spots in aged heart suggests a link between TFAM binding to mtDNA and loss of mitochondrial genome likely through hindrance of repair processes.

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