Strikingly higher frequency in centenarians and twins of mtDNA mutation causing remodeling of replication origin in leukocytes - PubMed (original) (raw)

Strikingly higher frequency in centenarians and twins of mtDNA mutation causing remodeling of replication origin in leukocytes

Jin Zhang et al. Proc Natl Acad Sci U S A. 2003.

Abstract

The presence of a genetic component in longevity is well known. Here, the association of a mtDNA mutation with a prolonged life span in humans was investigated. Large-scale screening of the mtDNA main control region in leukocytes from subjects of an Italian population revealed a homoplasmic C150T transition near an origin of heavy mtDNA-strand synthesis in approximately 17% of 52 subjects 99-106 years old, but, in contrast, in only 3.4% of 117 younger individuals (P = 0.0035). Evidence was obtained for the contribution of somatic events, under probable nuclear genetic control, to the striking selective accumulation of the mutation in centenarians. In another study, among leukocyte mtDNA samples from 20 monozygotic and 18 dizygotic twins, 60-75 years old, 30% (P = 0.0007) and 22% (P = 0.011), respectively, of the individuals involved exhibited the homoplasmic C150T mutation. In a different system, i.e., in five human fibroblast longitudinal studies, convincing evidence for the aging-related somatic expansion of the C150T mutation, up to homoplasmy, was obtained. Most significantly, 5' end analysis of nascent heavy mtDNA strands consistently revealed a new replication origin at position 149, substituting for that at 151, only in C150T mutation-carrying samples of fibroblasts or immortalized lymphocytes. Considering the aging-related health risks that the centenarians have survived and the developmental risks of twin gestations, it is proposed that selection for a remodeled replication origin, inherited or somatically acquired, provides a survival advantage and underlies the observed high incidence of the C150T mutation in centenarians and twins.

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Figures

Figure 1

Figure 1

Positions of the tissue-specific aging-dependent somatic mutations identified in human mtDNA main control region (1, 2) and of the C150T transition. OH1 and OH2, primary and secondary origin of H-strand synthesis; LSP, promoter for transcription of L-strand and synthesis of primer for H-strand synthesis (4, 5); CSB1, CSB2, and CSB3, conserved sequence blocks 1, 2, and 3 (6). The positions of binding of mitochondrial transcription factor A (mtTFA; the densely hatched rectangle indicates a position of high-affinity binding) are shown (1). Blue arrows and numbers indicate fibroblast-specific mutations (1), red arrows and numbers indicate skeletal muscle-specific mutations (2), and the green arrow and number indicate the C150T transition.

Figure 2

Figure 2

Bar graph summarizing the age distribution and frequency of the C150T transition (as determined by primer extension analysis) in mtDNA from lympho-monocytes (A), granulocytes (B), and buffy coats (C) of differently aged individuals. The number above each bar indicates the age in years of the individual analyzed. Red bars, C150T mutation-carrying mtDNA; green bars, wild-type mtDNA.

Figure 3

Figure 3

(A) Detection of C150T transition by allele-specific termination of primer extension, by using the PCR-amplified Init-Tra-Rep fragment as a template, in fibroblast mtDNA from the single biopsies of 32 subjects, 20-week fetal [FW (1)] and 1–103 years old, and from the second biopsies of the longitudinal studies LS12–LS19. The bar graph summarizes the age distribution and frequency of the C150T transition. (B) Detection of the C150T transition in the fibroblast mtDNA from the first and second biopsies of the longitudinal studies LS1–LS11, and age distribution and frequency of the mutation. W and M, primer extension products obtained by using as a template plasmid DNA carrying a cloned Init-Tra-Rep mtDNA fragment with wild-type and, respectively, mutant sequence; PE, primer+ sequenase only.

Figure 4

Figure 4

Detection of nascent H-mtDNA chains and identification of their 5′ ends. (A) Primer extension products of all nascent H-strand chains, obtained by using as templates the original mtDNA from the wild-type cell line AL4.27 or its PCR-amplified ACCI 2,818-bp fragment containing the mtDNA main control region (a), or the mtDNA from three fibroblast samples lacking (WT) and three fibroblast samples carrying (M) the C150T transition (b). (B) Primer extension products of nascent H-strand chains obtained by using as templates the mtDNA from AL4.27 cells (Ctrl), or from circulating granulocytes of centenarians lacking (WT) or carrying (M) the C150T transition (a), or from immortalized lymphocytes derived from centenarians lacking or carrying the mutation (b). In A and B, the age of the donors of fibroblasts, granulocytes, and immortalized lymphocytes is indicated above each lane. The 5′ ends of the nascent H-strand chains starting at positions 191, 167, 151, 149, and 146 were determined by sequencing, whereas the 5′ end of the 110 nascent chains was estimated from comparison of their migration with that of the marker (MSP1-digested pBR322 DNA).

Figure 5

Figure 5

Detection of the C150T transition by allele-specific termination of primer extension and its distribution and abundance in buffy coat mtDNA from MZ (A) and DZ (B) twins. Abbreviations are as in Fig. 3.

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