Multiple mechanisms for overcoming lethal over‐initiation of DNA replication
DNA replication is highly regulated and primarily controlled at the step of initiation. In bacteria, the replication initiator DnaA and the origin of replication oriC are the primary targets of regulation. Perturbations that increase or decrease replication initiation can cause a decrease in cell fi...
| Main Authors: | , , |
|---|---|
| Other Authors: | |
| Format: | Article |
| Language: | English |
| Published: |
Wiley
2022
|
| Online Access: | https://hdl.handle.net/1721.1/146802 |
| _version_ | 1811084766024302592 |
|---|---|
| author | Anderson, Mary E Smith, Janet L Grossman, Alan D |
| author2 | Massachusetts Institute of Technology. Department of Biology |
| author_facet | Massachusetts Institute of Technology. Department of Biology Anderson, Mary E Smith, Janet L Grossman, Alan D |
| author_sort | Anderson, Mary E |
| collection | MIT |
| description | DNA replication is highly regulated and primarily controlled at the step of initiation. In bacteria, the replication initiator DnaA and the origin of replication oriC are the primary targets of regulation. Perturbations that increase or decrease replication initiation can cause a decrease in cell fitness. We found that multiple mechanisms, including an increase in replication elongation and a decrease in replication initiation, can compensate for lethal over-initiation. We found that in Bacillus subtilis, under conditions of rapid growth, loss of yabA, a negative regulator of replication initiation, caused a synthetic lethal phenotype when combined with the dnaA1 mutation that also causes replication over-initiation. We isolated several classes of suppressors that restored viability to dnaA1 ∆yabA double mutants. Some suppressors (relA, nrdR) stimulated replication elongation. Others (dnaC, cshA) caused a decrease in replication initiation. One class of suppressors decreased replication initiation in the dnaA1 ∆yabA mutant by causing a decrease in the amount of the replicative helicase, DnaC. We found that decreased levels of helicase in otherwise wild-type cells were sufficient to decrease replication initiation during rapid growth, indicating that the replicative helicase is limiting for replication initiation. Our results highlight the multiple mechanisms cells use to regulate DNA replication. |
| first_indexed | 2024-09-23T12:57:14Z |
| format | Article |
| id | mit-1721.1/146802 |
| institution | Massachusetts Institute of Technology |
| language | English |
| last_indexed | 2024-09-23T12:57:14Z |
| publishDate | 2022 |
| publisher | Wiley |
| record_format | dspace |
| spelling | mit-1721.1/1468022022-12-09T03:22:01Z Multiple mechanisms for overcoming lethal over‐initiation of DNA replication Anderson, Mary E Smith, Janet L Grossman, Alan D Massachusetts Institute of Technology. Department of Biology DNA replication is highly regulated and primarily controlled at the step of initiation. In bacteria, the replication initiator DnaA and the origin of replication oriC are the primary targets of regulation. Perturbations that increase or decrease replication initiation can cause a decrease in cell fitness. We found that multiple mechanisms, including an increase in replication elongation and a decrease in replication initiation, can compensate for lethal over-initiation. We found that in Bacillus subtilis, under conditions of rapid growth, loss of yabA, a negative regulator of replication initiation, caused a synthetic lethal phenotype when combined with the dnaA1 mutation that also causes replication over-initiation. We isolated several classes of suppressors that restored viability to dnaA1 ∆yabA double mutants. Some suppressors (relA, nrdR) stimulated replication elongation. Others (dnaC, cshA) caused a decrease in replication initiation. One class of suppressors decreased replication initiation in the dnaA1 ∆yabA mutant by causing a decrease in the amount of the replicative helicase, DnaC. We found that decreased levels of helicase in otherwise wild-type cells were sufficient to decrease replication initiation during rapid growth, indicating that the replicative helicase is limiting for replication initiation. Our results highlight the multiple mechanisms cells use to regulate DNA replication. 2022-12-08T13:50:22Z 2022-12-08T13:50:22Z 2022 2022-12-08T13:43:56Z Article http://purl.org/eprint/type/JournalArticle https://hdl.handle.net/1721.1/146802 Anderson, Mary E, Smith, Janet L and Grossman, Alan D. 2022. "Multiple mechanisms for overcoming lethal over‐initiation of DNA replication." Molecular Microbiology, 118 (4). en 10.1111/MMI.14976 Molecular Microbiology Creative Commons Attribution NonCommercial License 4.0 https://creativecommons.org/licenses/by-nc/4.0/ application/pdf Wiley Wiley |
| spellingShingle | Anderson, Mary E Smith, Janet L Grossman, Alan D Multiple mechanisms for overcoming lethal over‐initiation of DNA replication |
| title | Multiple mechanisms for overcoming lethal over‐initiation of DNA replication |
| title_full | Multiple mechanisms for overcoming lethal over‐initiation of DNA replication |
| title_fullStr | Multiple mechanisms for overcoming lethal over‐initiation of DNA replication |
| title_full_unstemmed | Multiple mechanisms for overcoming lethal over‐initiation of DNA replication |
| title_short | Multiple mechanisms for overcoming lethal over‐initiation of DNA replication |
| title_sort | multiple mechanisms for overcoming lethal over initiation of dna replication |
| url | https://hdl.handle.net/1721.1/146802 |
| work_keys_str_mv | AT andersonmarye multiplemechanismsforovercominglethaloverinitiationofdnareplication AT smithjanetl multiplemechanismsforovercominglethaloverinitiationofdnareplication AT grossmanaland multiplemechanismsforovercominglethaloverinitiationofdnareplication |