Disintegration promotes protospacer integration by the Cas1-Cas2 complex
‘Disintegration’—the reversal of transposon DNA integration at a target site—is regarded as an abortive off-pathway reaction. Here, we challenge this view with a biochemical investigation of the mechanism of protospacer insertion, which is mechanistically analogous to DNA transposition, by the Strep...
| Main Authors: | , , , |
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| Format: | Article |
| Language: | English |
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eLife Sciences Publications Ltd
2021-08-01
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| Series: | eLife |
| Subjects: | |
| Online Access: | https://elifesciences.org/articles/65763 |
| _version_ | 1811201502733139968 |
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| author | Chien-Hui Ma Kamyab Javanmardi Ilya J Finkelstein Makkuni Jayaram |
| author_facet | Chien-Hui Ma Kamyab Javanmardi Ilya J Finkelstein Makkuni Jayaram |
| author_sort | Chien-Hui Ma |
| collection | DOAJ |
| description | ‘Disintegration’—the reversal of transposon DNA integration at a target site—is regarded as an abortive off-pathway reaction. Here, we challenge this view with a biochemical investigation of the mechanism of protospacer insertion, which is mechanistically analogous to DNA transposition, by the Streptococcus pyogenes Cas1-Cas2 complex. In supercoiled target sites, the predominant outcome is the disintegration of one-ended insertions that fail to complete the second integration event. In linear target sites, one-ended insertions far outnumber complete protospacer insertions. The second insertion event is most often accompanied by the disintegration of the first, mediated either by the 3′-hydroxyl exposed during integration or by water. One-ended integration intermediates may mature into complete spacer insertions via DNA repair pathways that are also involved in transposon mobility. We propose that disintegration-promoted integration is functionally important in the adaptive phase of CRISPR-mediated bacterial immunity, and perhaps in other analogous transposition reactions. |
| first_indexed | 2024-04-12T02:22:44Z |
| format | Article |
| id | doaj.art-cd580f2309094e43a0ef69a4c522eacb |
| institution | Directory Open Access Journal |
| issn | 2050-084X |
| language | English |
| last_indexed | 2024-04-12T02:22:44Z |
| publishDate | 2021-08-01 |
| publisher | eLife Sciences Publications Ltd |
| record_format | Article |
| series | eLife |
| spelling | doaj.art-cd580f2309094e43a0ef69a4c522eacb2022-12-22T03:52:05ZengeLife Sciences Publications LtdeLife2050-084X2021-08-011010.7554/eLife.65763Disintegration promotes protospacer integration by the Cas1-Cas2 complexChien-Hui Ma0Kamyab Javanmardi1https://orcid.org/0000-0002-6449-6709Ilya J Finkelstein2https://orcid.org/0000-0002-9371-2431Makkuni Jayaram3https://orcid.org/0000-0001-9640-6264Department of Molecular Biosciences and Institute of Cell and Molecular Biology, University of Texas at Austin, Austin, United StatesDepartment of Molecular Biosciences and Institute of Cell and Molecular Biology, University of Texas at Austin, Austin, United StatesDepartment of Molecular Biosciences and Institute of Cell and Molecular Biology, University of Texas at Austin, Austin, United States; Center for Systems and Synthetic Biology, University of Texas at Austin, Austin, United StatesDepartment of Molecular Biosciences and Institute of Cell and Molecular Biology, University of Texas at Austin, Austin, United States‘Disintegration’—the reversal of transposon DNA integration at a target site—is regarded as an abortive off-pathway reaction. Here, we challenge this view with a biochemical investigation of the mechanism of protospacer insertion, which is mechanistically analogous to DNA transposition, by the Streptococcus pyogenes Cas1-Cas2 complex. In supercoiled target sites, the predominant outcome is the disintegration of one-ended insertions that fail to complete the second integration event. In linear target sites, one-ended insertions far outnumber complete protospacer insertions. The second insertion event is most often accompanied by the disintegration of the first, mediated either by the 3′-hydroxyl exposed during integration or by water. One-ended integration intermediates may mature into complete spacer insertions via DNA repair pathways that are also involved in transposon mobility. We propose that disintegration-promoted integration is functionally important in the adaptive phase of CRISPR-mediated bacterial immunity, and perhaps in other analogous transposition reactions.https://elifesciences.org/articles/65763CRISPRadaptationtransposition |
| spellingShingle | Chien-Hui Ma Kamyab Javanmardi Ilya J Finkelstein Makkuni Jayaram Disintegration promotes protospacer integration by the Cas1-Cas2 complex eLife CRISPR adaptation transposition |
| title | Disintegration promotes protospacer integration by the Cas1-Cas2 complex |
| title_full | Disintegration promotes protospacer integration by the Cas1-Cas2 complex |
| title_fullStr | Disintegration promotes protospacer integration by the Cas1-Cas2 complex |
| title_full_unstemmed | Disintegration promotes protospacer integration by the Cas1-Cas2 complex |
| title_short | Disintegration promotes protospacer integration by the Cas1-Cas2 complex |
| title_sort | disintegration promotes protospacer integration by the cas1 cas2 complex |
| topic | CRISPR adaptation transposition |
| url | https://elifesciences.org/articles/65763 |
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