「様式３－B」
課題番号（Project number） 58

Characterization of a novel “DNA Maintenance Complex”(DMC) for genome integrity［１］組織（Research group）
代表者 (Principal Investigator)：
Lene Juel Rasmussen(University of Vopenhagen)
対応者 (Host researcher at IDAC)：
Akira Yasui(IDAC, Tohoku University)
研究費 (Expenditure report of research funds)：
consumable goods 100,000 Yen［２］研究経過（Research setup）
The goal of the proposed study is to demonstratethat mitochondrial dysfunction affects nuclear DNA
damage responses and thereby significantly
increases the risk of genomic instability and cellularsenescence.
Mitochondria houses rate-limiting enzymes
essential for de novo synthesis of pyrimidines. Incase of dysfunction of the mitochondrial respiration,
the cytosolic levels of especially pyrimidines, but alsopurines in the form of deoxyribonucleotide
phosphates (dNTPs) are decreased. We hypothesizethat this decrease will prevent the formation of a
proposed DNA maintenance complex (DMC) and inturn constrict the cells ability to perform nuclear
DNA bypass repair. As a result, DNA damages will
be sought salvaged by recombination repair, and therisk of chromosomal instability will increase.
Aim 1: To identify and characterize the DMC fromhuman cells in respect to protein components,
interaction with DNA and functionality at sites withDNA damage (Akaira Yasui, IDAC).
Aim 2: To demonstrate that dysfunction of themitochondrial respiration, in human cells constricts
synthesis of cellular dNTP, which in turn propagatesa signal comparable to a cellular starvation signal,

which prevents the formation of the DMC and as aresult restricts the cells ability to perform DNA
bypass repair (Lene Juel Rasmussen, UCPH).
［３］成果（Research outcomes）（３－１）研究成果（Results）
We have established a preliminary DMC
containing REV1, SHMT1, SHMT2, and SKAR2.
The preliminary results are based on proteomic
analysis of proteins interacting with known DMC
proteins such as Rev1. Fig. 1 depicts the results of anaffinity chromatography by using GST-tagged
SHMT1 purified from insect cells and massspectrometry to determine interacting proteins. The
results are very promising and the experimental
strategy will be continued.

Fig. 1M

BcGST

BcGST‐SHMT1kDa
CAD
220
160
120100

DHX9, ARMC5HNRNPU, SKAP2
NCL, DDX21
SHMT1, HSPA6KRT1050

BcGST‐M BcGST SHMT1302010（ ３ － ２ ） 波 及 効 果 と 発 展 性 な ど （ Futureperspectives）
If mitochondrial dysfunction is linked to dNTP
anabolism via the DMC, it could have implications
for understanding many aspects of human aging
and age-associated human disease. The results ofthe proposed studies will also contribute to basic
understanding of DNA repair, carcinogenesis,
nucleotide
metabolism,
andmitochondrial
dysfunction. In the long term, the proposed studies
could also stimulate development of novel diagnostic
and therapeutic agents for treating human disease.

［４］成果資料（List of Papers）
1. Fakouri NB, Durhuus JA, Regnell CE, Angleys M,Desler C, Hasan- Olive MDM, Martín-Pardillos A,
Tsaalbi-Shtylik A, Thomsen K, Lauritzen M, Bohr
VA, de Wind N, Bergersen LH, Rasmussen LJ. 2017.
Rev1 contributes to proper mitochondrial function
via the PARP-NAD+-SIRT1-PGC1α axis. Sci Rep.
Oct 2;7(1):12480. doi: 10.1038/s41598-017-12662-3.
2. Martín-Pardillos A, Tsaalbi-Shtylik A, Chen S,Lazare S, van Os RP, Dethmers-Ausema A, Fakouri
NB, Bosshard M, Aprigliano R, van Loon B,
Salvatori DCF, Hashimoto K, Dingemanse-van derSpek C, Moriya M, Rasmussen LJ, de Haan G,
Raaijmakers MHG, de Wind N. 2017. Genomic andfunctional integrity of the hematopoietic system
requires tolerance of oxidative DNA lesions. Blood.
Sep 28;130(13):1523-1534. doi: 10.1182/blood-201701-764274.
3. Thomsen K, Sherazi N, Yokota T, Hasan-Olive M,Fakouri NB, Desler C, Regnell C, Rasmussen LJ,Dela F, Bergersen LH, Lauritzen M. Initial brain
aging: heterogeneity of mitochondrial size isassociated with respiratory dysfunction in cortex
and hippocampus. 2017. Neurobiol Aging. Aug 12.
pii: S0197-4580(17)30259-2.
doi: 10.1016/j.neurobiolaging.2017.08.004.
4. Strickertsson J, Desler C, Rasmussen LJ. 2017.
Bacterial infection increases risk of carcinogenesisby targeting mitochondria. Semin Cancer Biol. Jul
25. pii: S1044-579X(17)30192-X.
doi: 10.1016/j.semcancer.2017.07.003.
5. Liu D, Keijzers G, Rasmussen LJ. 2017. DNAmismatch repair and its many roles in eukaryoticcells. Mutat Res Rev. 773:174-187.
doi: 10.1016/j.mrrev.2017.07.001. Epub 2017 Jul 9.
6. Liu D, Frederiksen JH, Liberti SE, Lützen A,Keijzers G, Pena-Diaz J, Rasmussen LJ. 2017.
Human DNA polymerase delta double-mutant
D316A E318A interferes with DNA mismatch
repair in vitro. Nucleic Acids Res. Sep 19;
45(16):9427-9440. doi: 10.1093/nar/gkx611.
7. Rudolph C, Nielsen JE, Jensen KV, Liu D, PenaDiaz J, Rajpert-De Meyts E, Rasmussen LJ,Jørgensen A. 2017. Involvement of the DNA
mismatch repair system in the cisplatin sensitivityof testicular germ cell tumours. Cell Oncol.
40(4):341-355. doi: 10.1007/s13402-017-0326-8.
8. Nielsen SV, Stein A, Dinitzen AB, Papaleo E,Tatham MH, Poulsen EG, Kassem MM, Rasmussen
LJ, Lindorff-Larsen K, Hartmann-Petersen R. 2017.
Predicting the impact of Lynch syndrome-causing
missense mutations from structural calculations.
PLOS Genet. 13(4):e1006739.
doi: 10.1371/journal.pgen.1006739. eCollection 2017
Apr.
9. Vohra R, Guruban IS, Henriksen U, Bergersen
LH, Rasmussen LJ, Desler C, Skytt DM, Kolko M.
2017. Disturbed mitochondrial function restricts
glutamate uptake in the human Müller glia cell line,MIO-M1. Mitochondrion. S1567-7249 (17) 30035-1.
10. Desler C, Lillenes MS, Tønjum T, Rasmussen LJ.
2017. The role of mitochondrial oxidative
phosphorylation in Alzheimer’s disease. Curr MedChem. doi: 10.2174/0929867324666170616110111.
11. Hopkinson BM, Desler C, Kalisz M, Vestentoft
PS, Rasmussen LJ, Bisgaard HC. 2017.
Bioenergetic Changes during Differentiation of
Human Embryonic Stem Cells along the HepaticLineage. Oxid Med Cell Longev. 2017;2017:5080128.
doi: 10.1155/2017/5080128.
12. Sykora P, Kanno S, Akbari M, Kulikowicz T,
Baptiste BA, Leandro G, Lu H, Tian J, May
A, Becker KA, Croteau DL, Wilson DM III, Sobol
RW, Yasui A and Bohr VA. DNA polymerase beta
participates in mitochondrial DNA repair. Mol CellBiol. 2017, doi: 10.1128/MCB.00237-17.