減數分裂遺傳重組與重複誘發點突變的分子機制

減數分裂是孟德爾遺傳學的核心。減數分裂是真核生物有性生殖細胞專有的細胞分裂方式，會使得染色體數目減半，製造出單倍體的精子、卵或真菌有性孢子。 我們實驗室以麵包酵母菌與工業用瑞氏木黴菌為模式，利用遺傳學、分子生物學、生物化學、結構生物學、基因體學與生物資訊學等方法，探討減數分裂細胞中 DNA 修補、同源重組與 DNA 損傷反應如何產生遺傳變異與維持基因體完整性。

重複誘發點突變（repeat-induced point mutation）是表觀遺傳學（epigenetics）的典範。重複誘發點突變是許多絲狀子囊菌（如工業用瑞氏木黴菌）有性生殖細胞特有的基因體防禦機制，利用兩個甲基化轉移酵素家族基因 dim2 與 rid1，將重複 DNA 序列與跳躍基因去活或滅能。

我們最近的研究可分為四大項：

1. 大多數進行有性生殖的真核生物（如酵母菌、擔子菌、果蠅、線蟲、高等植物與哺乳類）的減數分裂細胞，利用 Spo11 蛋白產生 DNA 雙股斷裂，從而啟動遺傳重組反應。我們發現剔除 spo11 基因的木黴菌仍能正常進行減數分裂與產生有性孢子。 目前研究重點在探討木黴菌如何啟動減數分裂遺傳重組反應。
2. 麵包酵母菌或人類均以Rad51 與Dmc1 兩個同源重組酵素催化減數分裂遺傳重組，木黴菌只有Rad51 無Dmc1。我們致力於比較分析麵包酵母菌Rad51、麵包酵母菌Dmc1 與木黴菌Rad51 的功能異同處。
3. DNA 損傷反應調控減數分裂遺傳重組、同源染色體聯會（synapsis）與染色體動態變化的分子機制。
4. 木黴菌的dim2 與rid1 將重複DNA 序列的胞嘧啶（C）先甲基化，再點突變成胸腺嘧啶（T）的化學機制。

• Education
• Selected publications
• Patents
• Lab members

• PDF, 1999-2000, Dept. Mol. & Cell Biol., Harvard Univ. USA
• Ph.D., 1998, Dept. Mol. & Cell Biol., Harvard Univ. USA
• MS, 1990, Inst. Life Science., Natl. Tsing Hua Univ.
• BS, 1988, Dept. Chemistry, Natl. Taiwan Univ.

1. Chen, C.L, Li, W.C., Chuang, Y.C., Liu, H.C., Huang, C.H., Lo, K.Y., Chen, C.Y., Chang, F.M., Chang, G. A., Lin, Y.L., Yang, W.D., Su, C.H., Yeh, T.M. and *Wang T.-F. (2022) Sexual crossing, chromosome-level genome sequences, and comparative genomic analyses for the medicinal mushroom Taiwanofungus camphoratus (Syn. Antrodia cinnamomea, Antrodia camphorata). Microbiology Spectrum 10, e02032-21. PMID:35196909
2. Li, W.C., Lin, T.C., Chen, C.C., Liu, H.C., Lin, H.N., Chao, J.L., Hsieh, C.H., Ni, H.F., Ruey- Chen, R.S.* and *Wang T.-F. (2021) Complete genome sequences and genome-wide characterization of Trichoderma biocontrol agents provide new insights into their evolution and variation in genome organization, sexual development, and fungal-plant interactions. Microbiology Spectrum 9, e00663-21. PMID:34908505.
3. Li, W.-C. #, Lee C.-Y. #, Lan, W.-H. #, Woo, T.-T. #, Liu, H.-C., Yeh, H.-Y., Chang, H.-Y., Chuang, Y.-C., Chen, C.-Y., Chuang, C.-N., Chen, C.-L., Hsueh, Y.-P., Li, *W.-H., P, *Chi., and *Wang T.-F. (2021) Trichoderma reesei Rad51 tolerates mismatches in hybrid meiosis with diverse genome sequences. Proc. Natl. Acad. Sci. USA 118. PMID: 33593897.
4. Liu, H.-C. #, #Li, W.-C.#, and *Wang, T.-F.* (2020) TSETA: a third-generation-sequencing based program for single-nucleotide-resolution mapping of meiotic recombination products and repeat-induced point mutations. Methods in Molecular Biology 2234: 331-361. PMID: 3316579.
5. Woo, T.T., Chuang, C.N., Higashide, M., Shinohara, A., *Wang, T.F. (2020) Dual roles of yeast Rad51 N-terminal domain in repairing DNA double-strand breaks. Nucleic Acids Research 48, 8474-8489 PMID:32652040.
6. Li, W.-C., Huang, C.-H., Chen, C.-L., Chuang, Y.-C., Tung, S.-Y. and *Wang T.-F. (2017) Trichoderma reesei complete genome sequence, repeat-induced point mutation and partitioning of CAZyme gene clusters. Biotechnology for Biofuels 10:170 PMID:28690679
7. Chen, Y.-J., Chuang, Y.-C., Chuang, C.-N., Cheng, Y.-H., Chang, C.-R., *Leng, C.-H. and *Wang, T.-F. (2016) Mre11 recruits conjugated SUMO moieties to facilitate the assembly and function of the Mre11-Rad50-Xrs2 complex. Nucleic Acids Research 44, 2199-2213. PMID:2.6743002
8. Cheng, Y.-H., Chuang, C.-N., Shen, H.-J., Lin, F.-M. and *Wang, T.-F. (2013) Three different modes of Mec1/ATR and Tel1/ATM activation in budding yeast early meiosis. Molecular and Cellular Biology 33, 3365-3376. PMID: 23775120.
9. Chuang, C.-N., Cheng, Y.-H. and *Wang, T.-F. (2012) Mek1 stabilizes Hop1-Thr318 phosphorylation to promote interhomolog recombination and checkpoint responses during yeast meiosis. Nucleic Acids Research 40, 11416-11427. PMID:23047948.
10. Lai, Y.-J., Lin, F.-M., Chuang, M.-J., Shen, H.-J. and *Wang, T.-F. (2011) Genetic requirements and functions of phosphorylation of the yeast axial element protein Red1. Molecular and Cell Biology 31, 912-923
11. Lin, F.-M., Lai, Y.-J., Shen, H.-J., Cheng, Y.-H. and *Wang, T.-F. (2010) Yeast axial element protein Red1 binds SUMO chains to promote meiotic interhomolog recombination and chromosome synapsis. EMBO Journal 29, 586-596. PMID:19959993
12. Chen, L.-T., Ko, T.-P., Chang, Y.-C., Lin, K.-A., Chang, C.-S., *Wang, A. H.-J. and *Wang, T.-F. (2007) Crystal structure of the left-handed Archaeal RadA helical filament: Identification of a new functional motif for controlling quaternary structures and enzymatic activity of RecA family proteins. Nucleic Acids Research 35, 1781-1801. PMID:17329376.
13. Cheng, C.-H., Lo, Y.-H., Liang, S.-S., Ti, S.-C., Lin, F.-M., Yeh, C.-H., Huang, H.-Y. and *Wang, T.-F. (2006) SUMO modifications control assembly of synaptonemal complex and polycomplex in meiosis of Saccharomyces cerevisiae. Genes & Development 20, 1986-1992. PMID: 16847351.
14. Chen, Y.-K., Leng, C.-H., Olivares, H., Lee, M.-H., Chang, Y.-C., Kung, W.-M., Ti., S.-C., Lo, Y.-H., Wang., A. H.-J., Chang, C.-S., Bishop, D. K., Hsueh, Y.-P. and *Wang, T.-F. (2004) Heterodimeric complexes of meiosis-specific Hop2 and Mnd1 promote DNA recombination and homolog juxtaposition. Proc. Natl. Acad. Sci. USA 101, 10572-10577. PMID: 15249670.

• Wang, T.-F., Chin-Ning Chuang and Tai-Ting Woo. “Method and vector for enhancing protein expression”. US provisional application (63/036,704), Patent Cooperation Treaty application (PCT/US2021/036247) and Taiwan application (110120541).
• Wang, T.-F., “Method for producing segmental aneuploidy (SAN) strains of Trichoderma reesei via sexual crossing and SAN strains produced therefrom”. Patent Cooperation Treaty (PCT/US14/58654) and US (9598738B2), Taiwan (I570236) and People’s Republic of China (IA001/ACA081CN).
• Wang, T. F. and Yang, S.M. “Methods of producing virus-like particles of piconavirus using a small-ubiquitin-related fusion protein expression system”. US (8663951B2) and Taiwan (I415623).
• Wang, T.-F. “SUMO fusion protein expression system for producing native proteins”. USA 8034910BZ.