Direct quantification of the translocation activities of Saccharomyces cerevisiae Pif1 helicase
Saccharomyces cerevisiae Pif1 (ScPif1) is known as an ATP-dependent DNA helicase that plays critical roles in a number of important biological processes such as DNA replication, telomere maintenance and genome stability maintenance. Besides its DNA helicase activity, ScPif1 is also known as a single...
| Main Authors: | , , , , , , |
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| Format: | Journal Article |
| Language: | English |
| Published: |
2020
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| Online Access: | https://hdl.handle.net/10356/142460 |
| _version_ | 1824456100368351232 |
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| author | Lu, Chen Le, Shimin Chen, Jin Byrd, Alicia K. Rhodes, Daniela Raney, Kevin D. Yan, Jie |
| author2 | School of Biological Sciences |
| author_facet | School of Biological Sciences Lu, Chen Le, Shimin Chen, Jin Byrd, Alicia K. Rhodes, Daniela Raney, Kevin D. Yan, Jie |
| author_sort | Lu, Chen |
| collection | NTU |
| description | Saccharomyces cerevisiae Pif1 (ScPif1) is known as an ATP-dependent DNA helicase that plays critical roles in a number of important biological processes such as DNA replication, telomere maintenance and genome stability maintenance. Besides its DNA helicase activity, ScPif1 is also known as a single-stranded DNA (ssDNA) translocase, while how ScPif1 translocates on ssDNA is unclear. Here, by measuring the translocation activity of individual ScPif1 molecules on ssDNA extended by mechanical force, we identified two distinct types of ssDNA translocation. In one type, ScPif1 moves along the ssDNA track with a rate of ∼140 nt/s in 100 μM ATP, whereas in the other type, ScPif1 is immobilized to a fixed location of ssDNA and generates ssDNA loops against force. Between the two, the mobile translocation is the major form at nanomolar ScPif1 concentrations although patrolling becomes more frequent at micromolar concentrations. Together, our results suggest that ScPif1 translocates on extended ssDNA in two distinct modes, primarily in a ‘mobile’ manner. |
| first_indexed | 2025-02-19T03:48:44Z |
| format | Journal Article |
| id | ntu-10356/142460 |
| institution | Nanyang Technological University |
| language | English |
| last_indexed | 2025-02-19T03:48:44Z |
| publishDate | 2020 |
| record_format | dspace |
| spelling | ntu-10356/1424602023-02-28T17:06:50Z Direct quantification of the translocation activities of Saccharomyces cerevisiae Pif1 helicase Lu, Chen Le, Shimin Chen, Jin Byrd, Alicia K. Rhodes, Daniela Raney, Kevin D. Yan, Jie School of Biological Sciences Science::Biological sciences Saccharomyces Cerevisiae Pif1 (ScPif1) ATP-dependent DNA Helicase Saccharomyces cerevisiae Pif1 (ScPif1) is known as an ATP-dependent DNA helicase that plays critical roles in a number of important biological processes such as DNA replication, telomere maintenance and genome stability maintenance. Besides its DNA helicase activity, ScPif1 is also known as a single-stranded DNA (ssDNA) translocase, while how ScPif1 translocates on ssDNA is unclear. Here, by measuring the translocation activity of individual ScPif1 molecules on ssDNA extended by mechanical force, we identified two distinct types of ssDNA translocation. In one type, ScPif1 moves along the ssDNA track with a rate of ∼140 nt/s in 100 μM ATP, whereas in the other type, ScPif1 is immobilized to a fixed location of ssDNA and generates ssDNA loops against force. Between the two, the mobile translocation is the major form at nanomolar ScPif1 concentrations although patrolling becomes more frequent at micromolar concentrations. Together, our results suggest that ScPif1 translocates on extended ssDNA in two distinct modes, primarily in a ‘mobile’ manner. MOE (Min. of Education, S’pore) Published version 2020-06-22T07:18:50Z 2020-06-22T07:18:50Z 2019 Journal Article Lu, C., Le, S., Chen, J., Byrd, A. K., Rhodes, D., Raney, K. D., & Yan, J. (2019). Direct quantification of the translocation activities of Saccharomyces cerevisiae Pif1 helicase. Nucleic Acids Research, 47(14), 7494-7501. doi:10.1093/nar/gkz541 0305-1048 https://hdl.handle.net/10356/142460 10.1093/nar/gkz541 31216020 2-s2.0-85071701208 14 47 7494 7501 en Nucleic Acids Research © 2019 The Author(s). Published by Oxford University Press on behalf of Nucleic Acids Research. This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/4.0/), which permits non-commercial re-use, distribution, and reproduction in any medium, provided the original work is properly cited. For commercial re-use, please contact journals.permissions@oup.com application/pdf |
| spellingShingle | Science::Biological sciences Saccharomyces Cerevisiae Pif1 (ScPif1) ATP-dependent DNA Helicase Lu, Chen Le, Shimin Chen, Jin Byrd, Alicia K. Rhodes, Daniela Raney, Kevin D. Yan, Jie Direct quantification of the translocation activities of Saccharomyces cerevisiae Pif1 helicase |
| title | Direct quantification of the translocation activities of Saccharomyces cerevisiae Pif1 helicase |
| title_full | Direct quantification of the translocation activities of Saccharomyces cerevisiae Pif1 helicase |
| title_fullStr | Direct quantification of the translocation activities of Saccharomyces cerevisiae Pif1 helicase |
| title_full_unstemmed | Direct quantification of the translocation activities of Saccharomyces cerevisiae Pif1 helicase |
| title_short | Direct quantification of the translocation activities of Saccharomyces cerevisiae Pif1 helicase |
| title_sort | direct quantification of the translocation activities of saccharomyces cerevisiae pif1 helicase |
| topic | Science::Biological sciences Saccharomyces Cerevisiae Pif1 (ScPif1) ATP-dependent DNA Helicase |
| url | https://hdl.handle.net/10356/142460 |
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