【14th.June】Nucleotide Excision Repair In Vivo:DNA Damage and Repair Maps of the Human Genome
日期：2017-06-14
閱讀：1504
TOPIC：Nucleotide Excision Repair In Vivo:DNA Damage and Repair Maps of the Human Genome
SPEAKER：胡晉川，研究助理，北卡大學醫(yī)學院生物化學與生物物理系，美國北卡羅萊納州，教堂山 27514,
TIME：June 14 (Wednesday)15:00pm?
LOCATION：化學B樓 410會議室
INVITER：仲冬平 教授
報告人簡介：
? ?胡晉川,研究助理,北卡大學醫(yī)學院生物化學與生物物理系,美國北卡羅萊納州，教堂山 27514,?
教育經歷
2004-2012 ? ? 理學博士，生物化學及分子生物學專業(yè),中國科學院微生物研究所?
研究經歷
2012.6- ? ? ? 博士后研究助理（-2017.5.31）/研究助理（2017.6.1-），北卡大學醫(yī)學院生化及生物物理系，導師：Aziz Sancar教授，美國科學院院士，2015年諾貝爾化學獎得主
研究內容：
? ?主要研究領域是核苷酸切除修復(Nucleotide Excision Repair, NER)，它可以修復環(huán)境致癌因子如紫外線, 以及化療藥物如順鉑等導致的DNA損傷，與癌癥發(fā)生和化療效果密切相關。我首次從細胞內分離了修復過程中切下的含有損傷的DNA片段（JBC 2013），這一發(fā)現(xiàn)是NER的切出機制（dual incision mechanism）最直接和確鑿的證據(jù)，而Sancar教授正是因為發(fā)現(xiàn)了NER的分子機制而獲得了2015年諾貝爾化學獎。在這個發(fā)現(xiàn)的基礎上，我建立了XR-seq的方法，通過對NER的切除片段進行高通量測序，第一次做到了在基因組水平上以單堿基分辨率來檢測NER（Gene Dev 2015）。利用該方法，我們發(fā)現(xiàn)基因組不同區(qū)域修復的先后順序受染色體結構的影響，且與癌癥的基因組的突變率相關（Adar, Hu et al. PNAS 2016）。除此之外，我還建立了Damage-seq，使我們第一次能在人類細胞中以單堿基分辨率檢測全基因組范圍的DNA加成物損傷的位置。用這種技術結合XR-seq，我獲得了抗癌藥物順鉑造成的DNA損傷與修復的圖譜（Hu et al. PNAS 2016）。在此基礎上我進一步提高了Damage-seq的靈敏度與特異性（HS-Damage-seq），檢測了人類細胞中UV損傷在形成和修復過程中的分布，發(fā)現(xiàn)了轉錄因子結合對DNA損傷形成的影響（PNAS 2017，已接收）。這兩種方法使我們第一次可以從基因組水平研究DNA損傷與修復，為從基因組水平上研究轉錄，染色體結構等因素對DNA損傷，修復及其引起的突變的影響，以及進一步對癌癥發(fā)生與化療效果等的影響提供了技術基礎。
2004.9-2012.4 博士研究生，中國科學院微生物研究所，導師：黃力研究員
博士期間我主要研究嗜高溫古菌硫磺礦硫化葉菌的DNA引發(fā)酶，發(fā)現(xiàn)該酶除了參與DNA復制，還可能參與了DNA雙鏈斷裂損傷修復的非同源末端連接途徑。
發(fā)表論文
1. Hu J*, Adebali O*, Adar S, and Sancar A. Dynamic maps of UV damage formation and repair for the human genome. Proc Natl Acad Sci U S A. 2017, In Press.
2. Hu J, Lieb JD, Sancar A, and Adar S. Cisplatin DNA damage and repair maps of the human genome at single-nucleotide resolution. Proc Natl Acad Sci U S A. 2016, 113(41):11507-11512 (Research Highlight In This Issue)
3. Adar S*, Hu J*, Lieb JD, and Sancar A. Genome-wide kinetics of DNA excision repair in relation to chromatin state and mutagenesis. Proc Natl Acad Sci U S A. 2016, 113(15): E2124-E2133
4. Hu J*, Adar S*, Selby CP, Lieb JD, and Sancar A. Genome-wide analysis of human global and transcription-coupled excision repair of UV damage at single-nucleotide resolution. Genes & Development. 2015, 29(9): 948-960. (Research Highlight In This Issue)
5. Hu J, Choi JH, Gaddameedhi S, Kemp MG, Reardon JT, Sancar A. Nucleotide excision repair in human cells: fate of the excised oligonucleotide carrying DNA damage in vivo. J Biol Chem. 2013, 288(29): 20918–20926.?
6. Hu J, Guo L, Wu K, Liu B, Lang S, Huang L. Template-dependent polymerization across discontinuous templates by the heterodimeric primase from the hyperthermophilic archaeon Sulfolobus solfataricus. Nucleic Acids Res. 2012, 40(8): 3470-3483
7. Hu J, Adar S. The Cartography of UV-induced DNA Damage Formation and DNA Repair. Photochem Photobiol. 2017, 93 (1): 199-206 Review.
8. Li W, Hu J, Adebali O, Adar S, Yang, Y, Chiou, YY, and Sancar A. General method for genome-wide mapping of nucleotide excision repair: application to UV and benzo[a]pyrene-induced DNA damage. Proc Natl Acad Sci U S A. 2017, In Press.
9. Adebali O, Chiou YY, Hu J, Sancar A, Selby CP. Genome-wide Transcription-coupled Repair in E. coli is Mediated by Mfd Protein. Proc Natl Acad Sci U S A. 2017, 114(11): E2116–E2125.
10. Kemp MG and Hu J. Post-excision Events in Human Nucleotide Excision Repair. Photochem Photobiol. 2017, 93 (1): 178-191 Review.
11. Canturk F, Karaman M, Selby CP, Kemp MG, Kulaksiz-Erkmen G, Hu J, Li W, Lindsey-Boltz LA, and Sancar A. Nucleotide excision repair by dual incisions in plants. Proc Natl Acad Sci U S A. 2016, 113(17):4706-10
12. Lindsey-Boltz LA, Kemp MG, Hu J, Sancar A. Analysis of ribonucleotide removal from DNA by human nucleotide excision repair. J Biol Chem. 2015, 290(50): 29801-7.
13. Sancar A, Lindsey-Boltz LA, Gaddameedhi S, Selby CP, Ye R, Chiou YY, Kemp MG, Hu J, Lee JH, and Ozturk N. Circadian Clock, Cancer, and Chemotherapy. Biochemistry, 2015, 54(2): pp 110–123. Review
14. Kemp MG, Gaddameedhi S, Choi JH, Hu J and Sancr A. DNA Repair Synthesis and Ligation Affect the Processing of Excised Oligonucleotides Generated by Human Nucleotide Excision Repair. J Biol Chem. 2014, 289(38): 26574-26583.?
15. Choi JH, Gaddameedhi S, Kim SY, Hu J, Kemp MG, Sancar A. Highly specific and sensitive method for measuring nucleotide excision repair kinetics of ultraviolet photoproducts in human cells. Nucleic Acids Res. 2014, 42(4): e29.
16. Liu Y, Guo L, Guo R, Wong RL, Hernandez H, Hu J, Chu Y, Amster IJ, Whitman WB, Huang L. The Sac10b homolog in Methanococcus maripaludis binds DNA at specific sites. J. Bacteriol. 2009, 191(7): 2315-29.
17. Wu K, Lai X, Guo X, Hu J, Xiang X, Huang L, Interplay between primase and replication factor C in the hyperthermophilic archaeon Sulfolobus solfataricus. Mol Microbiol. 2007, 63(3):826-37.
參與會議
1. 2016，美國國立癌癥研究所染色體生物學研討會，展板，美國，馬里蘭州，貝塞斯達
2. 2016，高登DNA損傷，突變與癌癥研究會議，展板，美國，加州，文圖拉
3. 2016，高登DNA損傷，突變與癌癥研討會，展板與報告，美國，加州，文圖拉
4. 2015，發(fā)育，衰老，疾病與表觀遺傳學研討會，展板，美國，北卡州，教堂山
5. 2015，美國國立癌癥研究所染色體生物學研討會, 展板，美國，馬里蘭州，貝塞斯達
6. 2013，美國國立衛(wèi)生研究院DNA修復視頻會議年輕研究者報告（每年僅選出三個年輕研究者，報告視頻鏈接：http://videocast.nih.gov/Summary.asp?File=18017&bhcp=1）
7. 2011，中國-丹麥古菌生物學及生物技術研討會，展板，中國，武漢
獎勵與榮譽
? 2015，北卡大學Joseph S. Pagano杰出論文獎
? 2015，北卡大學博士后杰出研究獎（每年全校所有專業(yè)獲獎者總計10人）
? 2012，北京市高校優(yōu)秀畢業(yè)生
? 2012，中國科學院研究生院優(yōu)秀畢業(yè)生
? 2012，中國科學院院長獎學金優(yōu)秀獎
? 2012，中國科學院微生物所所長獎學金
? 2009，中國科學院研究生院優(yōu)秀學生
報告摘要
Nucleotide Excision Repair In Vivo:
DNA Damage and Repair Maps of the Human Genome
AbstractDNA bulky adducts such as UV-induced damage can interfere with DNA replication and transcription, lead to mutation and cell death, and finally cause cancer and other disease. Nucleotide excision repair is the sole mechanism to repair these damage in humans. In nucleotide excision repair the damaged base(s) are removed by concerted dual incision bracketing the lesion to generate oligonucleotides of 26-27 nt in human cells. Though this reaction was reconstituted and well-studied in vitro, whether it has the same mechanism in vivo and how it is affected by chromosomal factors were unclear.?
To answer these questions, I isolated the in vivo excised oligonucleotides for the first time and demonstrated the congruence of in vivo and in vitro data on nucleotide excision repair. Then I developed methods for detecting the site of DNA bulky adducts formation (Damage-seq) and repair (XR-seq) at single nucleotide resolution and have generated damage and repair maps for the entire human genome. In Damage-seq, the exact locations of DNA damage are determined by the arrested DNA polymerase at damage sites. In XR-seq, the excised oligonucleotides containing DNA damage are captured by TFIIH-IP and subjected to high-throughput sequencing. DNA damage and repair maps generated by these methods indicated that while damage formation is mainly determined by sequence context and essentially uniform throughout the genome, repair is affected by chromatin states, transcription and other factors. The combination of damage and repair maps provides a holistic perspective of bulky adduct formation and repair of the human genome and has potential applications in cancer prevention and chemotherapy.
