Multi-site-specific endonucleases and the initiation of homologous genetic recombination in yeast |
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| Institution: | 1. Institute of Biological and Medical Engineering, Guangdong Academy of Sciences, Guangzhou 510632, China;2. Hubei Province Research Center of Engineering Technology for Utilization of Botanical Functional Ingredients, Hubei Engineering University, China;3. Department of Orthopedic Surgery, The Second Affiliated Hospital of Guangzhou Medical University, Guangzhou 510632, China;4. Guangdong Provincial Key Lab of Medical Electronic Instruments and Polymer Material Products, Guangzhou 510632, China;5. Department of Orthopaedics, the First Affiliated Hospital of Nanchang University, 330006 Nanchang, China;1. Department of Urology, Frontier Science Center for Immunology and Metabolism, Medical Research Institute, Zhongnan Hospital of Wuhan University, Wuhan University, Wuhan 430071, China;2. Department of Pathology, Frontier Science Center for Immunology and Metabolism, Medical Research Institute, Zhongnan Hospital of Wuhan University, Wuhan University, Wuhan 430071, China;3. Medical Research Institute, Renmin Hospital of Wuhan University, Wuhan University, Wuhan, China;1. State Key Laboratory of Southwestern Chinese Medicine Resources, Pharmacy School, Chengdu University of Traditional Chinese Medicine, Chengdu 611130, China;2. TCM Regulating Metabolic Diseases Key Laboratory of Sichuan Province, Hospital of Chengdu University of Traditional Chinese Medicine, Chengdu 610072, China;3. Sichuan Provincial Acupuncture School, Chengdu 611731, China;4. Chengdu Huashen Technology Group Co., Ltd., Chengdu 611137, Sichuan, China |
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| Abstract: | The notion that homologous recombination is a regulated biological process is not a familiar one. In yeasts, homologous recombination and most site-specific ones are initiated by site-specific double-stranded breaks that are introduced within cis-acting elements for the recombination. On the other hand, yeasts have a group of site-specific endonucleases (multi-site-specific endonucleases) that have a number of cleavage sites on each DNA. One of them, Endo.SceI of S. cerevisiae, was shown to introduce double-stranded breaks at a number of welldefined sites on the mitochondrial DNA in vivo. An Endo.SceI-induced double-stranded break was demonstrated to induce homologous recombination in mitochondria. Like the case of homologous recombination of nuclear chromosomes, the double-stranded break induces gene conversion of both genetic markers flanking and in the proximity of the cleavage site, and the cleaved DNA acts as a recipient of genetic information from the uncleaved partner DNA. The 70 kDa-heat-shock protein (HSP70)-subunit of Endo.SceI and a general role of the HSP70 in the regulation of protein-folding suggest the regulation of nucleolytic activity of Endo.SceI. |
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