A protocol to determine the activities of human MUS81-EME1&2 endonucleases
Summary: The human MUS81-EME1&2 complexes are structure-selective endonucleases that play important roles in DNA damage repair. Here, we describe a protocol to determine the endonuclease activities of MUS81-EME1&2 complexes toward various DNA structures. We co-express MUS81 with EME1 or EME2...
| Main Authors: | , , |
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| Format: | Article |
| Language: | English |
| Published: |
Elsevier
2022-09-01
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| Series: | STAR Protocols |
| Subjects: | |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S2666166722004087 |
| _version_ | 1818532783760343040 |
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| author | Qianqian Fang Zhengkang Hua Zhonghui Lin |
| author_facet | Qianqian Fang Zhengkang Hua Zhonghui Lin |
| author_sort | Qianqian Fang |
| collection | DOAJ |
| description | Summary: The human MUS81-EME1&2 complexes are structure-selective endonucleases that play important roles in DNA damage repair. Here, we describe a protocol to determine the endonuclease activities of MUS81-EME1&2 complexes toward various DNA structures. We co-express MUS81 with EME1 or EME2 and purify the complexes with high purity, and determine their activities on the cleavages of 3ˊ flaps, 5ˊ flaps, nicked double-stranded DNAs, and Holliday junctions. This protocol can also be used for the determination of substrate preferences of other structure-selective endonucleases.For complete details on the use and execution of this protocol, please refer to Hua et al. (2022). : Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics. |
| first_indexed | 2024-12-11T17:50:02Z |
| format | Article |
| id | doaj.art-5418d93c1fd64e71a39bd5bc508188a2 |
| institution | Directory Open Access Journal |
| issn | 2666-1667 |
| language | English |
| last_indexed | 2024-12-11T17:50:02Z |
| publishDate | 2022-09-01 |
| publisher | Elsevier |
| record_format | Article |
| series | STAR Protocols |
| spelling | doaj.art-5418d93c1fd64e71a39bd5bc508188a22022-12-22T00:56:15ZengElsevierSTAR Protocols2666-16672022-09-0133101528A protocol to determine the activities of human MUS81-EME1&2 endonucleasesQianqian Fang0Zhengkang Hua1Zhonghui Lin2College of Chemistry, Fuzhou University, Fuzhou 350108, ChinaCollege of Chemistry, Fuzhou University, Fuzhou 350108, ChinaCollege of Chemistry, Fuzhou University, Fuzhou 350108, China; Corresponding authorSummary: The human MUS81-EME1&2 complexes are structure-selective endonucleases that play important roles in DNA damage repair. Here, we describe a protocol to determine the endonuclease activities of MUS81-EME1&2 complexes toward various DNA structures. We co-express MUS81 with EME1 or EME2 and purify the complexes with high purity, and determine their activities on the cleavages of 3ˊ flaps, 5ˊ flaps, nicked double-stranded DNAs, and Holliday junctions. This protocol can also be used for the determination of substrate preferences of other structure-selective endonucleases.For complete details on the use and execution of this protocol, please refer to Hua et al. (2022). : Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics.http://www.sciencedirect.com/science/article/pii/S2666166722004087Molecular BiologyProtein BiochemistryProtein expression and purificationStructural Biology |
| spellingShingle | Qianqian Fang Zhengkang Hua Zhonghui Lin A protocol to determine the activities of human MUS81-EME1&2 endonucleases STAR Protocols Molecular Biology Protein Biochemistry Protein expression and purification Structural Biology |
| title | A protocol to determine the activities of human MUS81-EME1&2 endonucleases |
| title_full | A protocol to determine the activities of human MUS81-EME1&2 endonucleases |
| title_fullStr | A protocol to determine the activities of human MUS81-EME1&2 endonucleases |
| title_full_unstemmed | A protocol to determine the activities of human MUS81-EME1&2 endonucleases |
| title_short | A protocol to determine the activities of human MUS81-EME1&2 endonucleases |
| title_sort | protocol to determine the activities of human mus81 eme1 2 endonucleases |
| topic | Molecular Biology Protein Biochemistry Protein expression and purification Structural Biology |
| url | http://www.sciencedirect.com/science/article/pii/S2666166722004087 |
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