Increasing the efficiency of HR (homologous recombination) is important for a successful knock-in. Rad51 is mainly involved in homologous recombination and is associated with strand invasion. The HR-related mismatch repair system maintains HR fidelity by heteroduplex rejection and repair. Therefore, the purpose of this study is to control Rad51, which plays a critical role in HR, through UV-induced DNA damage. It is also to confirm the effect on the expression of MMR related genes (Msh2, Msh3, Msh6, Mlh1, Pms2) and HR-related genes closely related to HR through treatment with the MMR inhibitor CdCl2. The mRNA expression of Rad51 gene was confirmed in both HC11 cells and mouse testes, but the mRNA expression of Dmc1 gene was confirmed only in mouse testes. The protein expression of Rad51 and Dmc1 gene increased in UV-irradiated HC11 cells. After 72 hours of treatment with 1 μm of CdCl2, the mRNA expression level of Msh3, Pms2, and Rad51 decreased, but the mRNA expression level of Msh6 and Mlh1 increased in HC11 cells. There was no significant difference in Msh2 mRNA expression between CdCl2 untreated-group and the 72 hours treated group. In conclusion, HR-related gene (Rad51) was increased by UV-induced DNA damage. Treatment of the MMR inhibitor CdCl2 in HC11 cells decreased the mRNA expression of Rad51.
The acetyltransferase Tip60 (Kat5) is a member of the MYST family of HATs that was initially identified as a cellular protein. TIP60 acetylates histone and non-histone proteins, and is involved in diverse biological processes, including apoptosis, cell cycle, and DNA damage responses. In this study, a specific inhibitor of TIP60, Nu 9056, was used to study the function and its regulatory mechanism of Tip60 in the porcine preimplantation embryonic development. The results showed that inhibition of TIP60 impaired the embryonic development due to induce DNA damage through ATM-p53-p21 pathway, it was evidenced by expression of γH2A in the nuclei of blastocysts. In addition, TIP60 inhibition decreased efficiency of DNA repair by regulating P53 binding protein 1 expression. Furthermore, autophagy was induced following TIP60 inhibition through modulating microtubule-associated protein 1A/1B-light chain 3 expression. In conclusion, the results suggest that TIP60 plays a critical role in early embryonic development via regulation of DNA damage and its repairs.
The effects of extracts from Lentinula edodes (L. edodes) and Aquilariae agallocha (A. agallocha) on the DNA damage response in ultraviolet A (UVA)-exposed HaCaT cells and on the allergic contact dermatitis caused by 2,4-dinitro-chlorobezene (DNCB) were investigated. When UVA-exposed cells were incubated for 24 hours in medium containing L. edodes or A. agallocha extract, the level of 8-OHdG and CPD decreased in a concentration-dependent manner. The combined treatment with both extracts potentiated the decrease in UVA-induced 8-OHdG and CPD levels as compared with those following treatment with a single extract. In addition, the two extracts showed preventive effects against the UVA-induced reduction in collagen levels. Furthermore, the blood levels of IgE, IL-6, and histamine decreased more significantly upon combined treatment with L. edodes and A. agallocha extracts as compared with those following treatment with single extracts in DNCB-induced allergic contact dermatitis in the ICR mouse. The results of the present study suggest that the components with in the extracts of L. edodes and A. agallocha can help to prevent of UVA-induced genomic instability via a decrease in DNA damage, and to decrease the DNCB-induced allergic dermatitis via modulation of relevant proteins including IgE and IL-6. Further study is needed to clarify the purified components related to the preventative effects of the two extracts against UVA- or DNCB-induced genomic damage.
수은이 DNA 수복에 미치는 영향을 알아보기 위해 E. fetida를 염화수은(II)과 이온화 방사선에 순차적으로 노출시킨 후, 단세포 겔 전기영동 기법을 이용하여 DNA의 손상 수준과 방사선 조사 후 시간 경과에 따른 수복 양상을 관찰하였다. 염화수은(II)의 농도를 40 mg kg-1으로 하여 48시간 동안 in vivo 노출 시험을 수행한 뒤 20Gy의 감마선을 조사한 결과, 시간이 지날수록 대체로 DNA 손상의 수준이 감소했다. 이온화
E. fetida를 방사선과 수은에 각각 노출시킨 후, 체강세포를 추출하고 단세포 겔 전기영동 기법을 이용하여 DNA의 손상정도와 시간의 경과에 따른 수복 양상을 평가해 보았다. 그 결과, 방사선 조사 후의 시간이 경과할수록 대체로 DNA 손상정도가 감소했으며, 12시간 내에 모든 실험군의 DNA가 완전히 수복되었다. 정확한 수복 완료 시간을 알아보기 위해 OTM 값을 대조군과 비교해 보면 2.5와 5Gy는 방사선 조사 후 약 2시간, 10과 20 Gy
Rad51 is a key component of homologous recombination (HR) to repair DNA double-strand breaks and it forms Rad51 recombinase filaments of broken single-stranded DNA to promote HR. In addition to its role in DNA repair and cell cycle progression, Rad51 contributes to the reprogramming process during the generation of induced pluripotent stem cells. In light of this, we performed reprogramming experiments to examine the effect of co-expression of Rad51 and four reprogramming factors, Oct4, Sox2, Klf4, and c-Myc, on the reprogramming efficiency. Co-expression of Rad51 significantly increased the numbers of alkaline phosphatase-positive colonies and embryonic stem cell-like colonies during the process of reprogramming. Co-expression ofRad51 significantly increased the expression of epithelial markers at an early stage of reprogramming compared with control cells. Phosphorylated histone H2AX (γH2AX), which initiates the DNA double-strand break repair system, was highly accumulated in reprogramming intermediates upon co-expression of Rad51. This study identified a novel role of Rad51 in enhancing the reprogramming efficiency, possibly by facilitating mesenchymal-to-epithelial transition and by regulating a DNA damage repair pathway during the early phase of the reprogramming process.
DNA 회복효소인 MPG는 DNA의 퓨린기에 결합되어 있는 메틸기 등 이물질을 염기와 함께 제거하는 작용을 한다. 본 연구에서는 노던 블롯팅 방법을 이용하여 Balb/c mice의 각 조직별로 발생단계별 mRNA 발현 정도를 조사하였다. 뇌와 콩팥조직에서는 출생직후에 발현이 가장 활발하였으며, 성체시기까지 비교적 높은 활성도가 유지되었다. 위장 조직에서는 출생직후에서 일주일 후까지는 명확히 관찰되었으나, 그 이후는 발현이 약화되었다. 간장과 폐조직에서는