X-Message-Number: 25665 Date: Sun, 6 Feb 2005 05:26:59 -0800 (PST) From: Doug Skrecky <> Subject: DNA polymerase beta rejuvenates DNA repair J Neurochem. 2005 Feb;92(4):818-23. Reduced DNA gap repair in aging rat neuronal extracts and its restoration by DNA polymerase beta and DNA-ligase. Abstract Synthetic deoxy-oligo duplexes containing short gaps of 1 and 4 nucleotides were used as model substrates to assess the DNA gap repair ability of the neuronal extracts prepared from cerebral cortex of rats of different ages. Our results demonstrate that gap repair activity in neurons decreases markedly with age. The decreased activity could be restored by supplementing the neuronal extracts with pure recombinant rat liver DNA polymerase beta. High levels of DNA polymerase beta supplementation resulted in gap-filling activity that proceeded essentially through addition of nucleotides through a slow distributive strand displacement mode to achieve full template length (32-mer). However, at lower concentrations of DNA polymerase beta, the gap repair takes place quickly through gap filling followed by ligation to downstream primer, in an energy efficient manner. For this to happen, the conditions required are the presence of 5'-PO(4) on the downstream primer and supplementation of aging neuronal extracts with DNA-ligase in addition to recombinant DNA polymerase beta. These results demonstrate that aging neurons are unable to affect base excision repair (BER) due to deficiency of DNA polymerase beta and DNA-ligase and fortifying aged neuronal extracts with these two factors can restore the lost BER activity. [Could this be how CR affects cancer risk?] DNA Repair (Amst). 2003 Mar 1;2(3):295-307 Caloric restriction promotes genomic stability by induction of base excision repair and reversal of its age-related decline. Caloric restriction is a potent experimental manipulation that extends mean and maximum life span and delays the onset and progression of tumors in laboratory rodents. While caloric restriction (CR) clearly protects the genome from deleterious damage, the mechanism by which genomic stability is achieved remains unclear. We provide evidence that CR promotes genomic stability by increasing DNA repair capacity, specifically base excision repair (BER). CR completely reverses the age-related d ecline in BER capacity (P<0.01) in all tissues tested (brain, liver, spleen and testes) providing aged, CR animals with the BER phenotype of young, ad libitum-fed animals. This CR-induced reversal of the aged BER phenotype is accompanied by a reversal in the age-related decline in DNA polymerase beta (beta-pol), a rate-limiting enzyme in the BER pathway. CR significantly reversed the age-related loss of beta-pol protein levels (P<0.01), mRNA levels (P<0.01) and enzyme activity (P<0.01) in all tissues tested. Additionally, in young (4-6-month-old) CR animals a significant up-regulation in BER capacity, beta-pol protein and beta-pol mRNA is observed (P<0.01), demonstrating an early effect of CR that may provide insight in distinguishing the anti-tumor from the anti-aging effects of CR. This up-regulation in BER by caloric restriction in young animals corresponds to increased protection from carcinogen exposure, as mutation frequency is significantly reduced in CR animals exposed to either DMS or 2-nitropropane (2-NP) (P<0.01). Overall the data suggest an important biological consequence of moderate BER up-regulation and provides support for the hormesis theory of caloric restriction. Mech Ageing Dev. 1998 Aug 14;104(2):133-48. Improved DNA-repair parameters in PHA-stimulated peripheral blood lymphocytes of human subjects with low body mass index. Clinically healthy subjects of the Indian population were divided into three age groups: young, 8-14 years; adult, 20-35 years; old, > or = 55 years and were further classified based on body mass index (BMI) as normal BMI (NBMI)> or =20 and low BMI (LBMI) between 16 and 18, respectively. The ability of the peripheral blood lymphocytes from these subjects to respond to PHA stimulation in vitro and DNA-repair parameters, thereafter as a function of BMI and aging, were studied. The DNA-repair markers like unscheduled DNA synthesis (UDS), activities of DNA polymerase beta and of two endodeoxy-ribonucleases, (UV- and AP-DNases) were assessed under different conditions. The LBMI group, considered to be going through chronic but mild undernutrition, showed higher repair capacity and exhibited no appreciable age-dependent decline in DNA-repair potential as was seen in normal subjects. These results correlate well with those seen in unstimulated human lymphocytes and also confirm the observations made earlier in experimental animals, where dietary restriction was shown to have beneficial effects on DNA-repair capacity. Rate This Message: http://www.cryonet.org/cgi-bin/rate.cgi?msg=25665