A CRISPR interference screen reveals a role for cell wall teichoic acids in conjugation in Bacillus subtilis
Conjugative elements are widespread in bacteria and include plasmids and integrative and conjugative elements (ICEs). They transfer from donor to recipient cells via an element-encoded type IV secretion system. These elements interact with and utilize host functions for their lifecycles. We sought t...
| Main Authors: | , , |
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| Format: | Article |
| Language: | English |
| Published: |
Wiley
2022
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| Online Access: | https://hdl.handle.net/1721.1/144143 |
| _version_ | 1811089148476391424 |
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| author | Harden, M Michael Anderson, Mary E Grossman, Alan D |
| author2 | Massachusetts Institute of Technology. Department of Biology |
| author_facet | Massachusetts Institute of Technology. Department of Biology Harden, M Michael Anderson, Mary E Grossman, Alan D |
| author_sort | Harden, M Michael |
| collection | MIT |
| description | Conjugative elements are widespread in bacteria and include plasmids and integrative and conjugative elements (ICEs). They transfer from donor to recipient cells via an element-encoded type IV secretion system. These elements interact with and utilize host functions for their lifecycles. We sought to identify essential host genes involved in the lifecycle of the integrative and conjugative element ICEBs1 of Bacillus subtilis. We constructed a library of strains for inducible knockdown of essential B. subtilis genes using CRISPR interference. Each strain expressed one guide RNA in ICEBs1. We induced partial interference of essential genes and identified those that caused an acute defect in acquisition of ICEBs1 by recipient cells. This screen revealed that reducing expression of genes needed for synthesis of cell wall teichoic acids caused a decrease in conjugation. Using three different ways to reduce their synthesis, we found that wall teichoic acids were necessary in both donors and recipients for efficient conjugative transfer of ICEBs1. Further, we found that depletion of wall teichoic acids caused cells involved in ICEBs1 conjugation to die, most likely from damage to the cell envelope. Our results indicate that wall teichoic acids help protect against envelope stress caused by active conjugation machines. |
| first_indexed | 2024-09-23T14:14:27Z |
| format | Article |
| id | mit-1721.1/144143 |
| institution | Massachusetts Institute of Technology |
| language | English |
| last_indexed | 2024-09-23T14:14:27Z |
| publishDate | 2022 |
| publisher | Wiley |
| record_format | dspace |
| spelling | mit-1721.1/1441432023-04-13T16:09:08Z A CRISPR interference screen reveals a role for cell wall teichoic acids in conjugation in Bacillus subtilis Harden, M Michael Anderson, Mary E Grossman, Alan D Massachusetts Institute of Technology. Department of Biology Conjugative elements are widespread in bacteria and include plasmids and integrative and conjugative elements (ICEs). They transfer from donor to recipient cells via an element-encoded type IV secretion system. These elements interact with and utilize host functions for their lifecycles. We sought to identify essential host genes involved in the lifecycle of the integrative and conjugative element ICEBs1 of Bacillus subtilis. We constructed a library of strains for inducible knockdown of essential B. subtilis genes using CRISPR interference. Each strain expressed one guide RNA in ICEBs1. We induced partial interference of essential genes and identified those that caused an acute defect in acquisition of ICEBs1 by recipient cells. This screen revealed that reducing expression of genes needed for synthesis of cell wall teichoic acids caused a decrease in conjugation. Using three different ways to reduce their synthesis, we found that wall teichoic acids were necessary in both donors and recipients for efficient conjugative transfer of ICEBs1. Further, we found that depletion of wall teichoic acids caused cells involved in ICEBs1 conjugation to die, most likely from damage to the cell envelope. Our results indicate that wall teichoic acids help protect against envelope stress caused by active conjugation machines. 2022-07-29T17:27:17Z 2022-07-29T17:27:17Z 2022-06 2022-07-29T17:21:03Z Article http://purl.org/eprint/type/JournalArticle https://hdl.handle.net/1721.1/144143 Harden, M Michael, Anderson, Mary E and Grossman, Alan D. 2022. "A CRISPR interference screen reveals a role for cell wall teichoic acids in conjugation in Bacillus subtilis." Molecular Microbiology, 117 (6). en 10.1111/mmi.14914 Molecular Microbiology Creative Commons Attribution-NonCommercial-NoDerivs License http://creativecommons.org/licenses/by-nc-nd/4.0/ application/pdf Wiley Wiley |
| spellingShingle | Harden, M Michael Anderson, Mary E Grossman, Alan D A CRISPR interference screen reveals a role for cell wall teichoic acids in conjugation in Bacillus subtilis |
| title | A CRISPR interference screen reveals a role for cell wall teichoic acids in conjugation in Bacillus subtilis |
| title_full | A CRISPR interference screen reveals a role for cell wall teichoic acids in conjugation in Bacillus subtilis |
| title_fullStr | A CRISPR interference screen reveals a role for cell wall teichoic acids in conjugation in Bacillus subtilis |
| title_full_unstemmed | A CRISPR interference screen reveals a role for cell wall teichoic acids in conjugation in Bacillus subtilis |
| title_short | A CRISPR interference screen reveals a role for cell wall teichoic acids in conjugation in Bacillus subtilis |
| title_sort | crispr interference screen reveals a role for cell wall teichoic acids in conjugation in bacillus subtilis |
| url | https://hdl.handle.net/1721.1/144143 |
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