Investigating the role of Rts1 in DNA replication initiation
<strong>Background:</strong> Understanding DNA replication initiation is essential to understand the mis-regulation of replication seen in cancer and other human disorders. DNA replication initiates from DNA replication origins. In eukaryotes, replication is dependent on cell cycle kinas...
| Main Authors: | , |
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| Format: | Journal article |
| Language: | English |
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F1000Research
2018
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| _version_ | 1797072877939326976 |
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| author | Wallis, A Nieduszynski, C |
| author_facet | Wallis, A Nieduszynski, C |
| author_sort | Wallis, A |
| collection | OXFORD |
| description | <strong>Background:</strong> Understanding DNA replication initiation is essential to understand the mis-regulation of replication seen in cancer and other human disorders. DNA replication initiates from DNA replication origins. In eukaryotes, replication is dependent on cell cycle kinases which function during S phase. Dbf4-dependent kinase (DDK) and cyclin-dependent kinase (CDK) act to phosphorylate the DNA helicase (composed of mini chromosome maintenance proteins: Mcm2-7) and firing factors to activate replication origins. It has recently been found that Rif1 can oppose DDK phosphorylation. Rif1 can recruit protein phosphatase 1 (PP1) to dephosphorylate MCM and restricts origin firing. In this study, we investigate a potential role for another phosphatase, protein phosphatase 2A (PP2A), in regulating DNA replication initiation. The PP2A regulatory subunit Rts1 was previously identified in a large-scale genomic screen to have a genetic interaction with ORC2 (a DNA replication licensing factor). Deletion of RTS1 synthetically rescued the temperature-sensitive (ts-) phenotype of ORC2 mutants. <strong>Methods:</strong> We deleted RTS1 in multiple ts-replication factor Saccharomyces cerevisiae strains, including ORC2. Dilution series assays were carried out to compare qualitatively the growth of double mutant ∆rts1 ts-replication factor strains relative to the respective single mutant strains. <strong>Results:</strong> No synthetic rescue of temperature-sensitivity was observed. Instead we found an additive phenotype, indicating gene products function in separate biological processes. These findings are in agreement with a recent genomic screen which found that RTS1 deletion in several ts-replication factor strains led to increased temperature-sensitivity. <strong>Conclusions:</strong> We find no evidence that Rts1 is involved in the dephosphorylation of DNA replication initiation factors. |
| first_indexed | 2024-03-06T23:14:01Z |
| format | Journal article |
| id | oxford-uuid:667ad7c1-148b-4f35-bae9-a9e0503d75c8 |
| institution | University of Oxford |
| language | English |
| last_indexed | 2024-03-06T23:14:01Z |
| publishDate | 2018 |
| publisher | F1000Research |
| record_format | dspace |
| spelling | oxford-uuid:667ad7c1-148b-4f35-bae9-a9e0503d75c82022-03-26T18:32:11ZInvestigating the role of Rts1 in DNA replication initiationJournal articlehttp://purl.org/coar/resource_type/c_dcae04bcuuid:667ad7c1-148b-4f35-bae9-a9e0503d75c8EnglishSymplectic Elements at OxfordF1000Research2018Wallis, ANieduszynski, C<strong>Background:</strong> Understanding DNA replication initiation is essential to understand the mis-regulation of replication seen in cancer and other human disorders. DNA replication initiates from DNA replication origins. In eukaryotes, replication is dependent on cell cycle kinases which function during S phase. Dbf4-dependent kinase (DDK) and cyclin-dependent kinase (CDK) act to phosphorylate the DNA helicase (composed of mini chromosome maintenance proteins: Mcm2-7) and firing factors to activate replication origins. It has recently been found that Rif1 can oppose DDK phosphorylation. Rif1 can recruit protein phosphatase 1 (PP1) to dephosphorylate MCM and restricts origin firing. In this study, we investigate a potential role for another phosphatase, protein phosphatase 2A (PP2A), in regulating DNA replication initiation. The PP2A regulatory subunit Rts1 was previously identified in a large-scale genomic screen to have a genetic interaction with ORC2 (a DNA replication licensing factor). Deletion of RTS1 synthetically rescued the temperature-sensitive (ts-) phenotype of ORC2 mutants. <strong>Methods:</strong> We deleted RTS1 in multiple ts-replication factor Saccharomyces cerevisiae strains, including ORC2. Dilution series assays were carried out to compare qualitatively the growth of double mutant ∆rts1 ts-replication factor strains relative to the respective single mutant strains. <strong>Results:</strong> No synthetic rescue of temperature-sensitivity was observed. Instead we found an additive phenotype, indicating gene products function in separate biological processes. These findings are in agreement with a recent genomic screen which found that RTS1 deletion in several ts-replication factor strains led to increased temperature-sensitivity. <strong>Conclusions:</strong> We find no evidence that Rts1 is involved in the dephosphorylation of DNA replication initiation factors. |
| spellingShingle | Wallis, A Nieduszynski, C Investigating the role of Rts1 in DNA replication initiation |
| title | Investigating the role of Rts1 in DNA replication initiation |
| title_full | Investigating the role of Rts1 in DNA replication initiation |
| title_fullStr | Investigating the role of Rts1 in DNA replication initiation |
| title_full_unstemmed | Investigating the role of Rts1 in DNA replication initiation |
| title_short | Investigating the role of Rts1 in DNA replication initiation |
| title_sort | investigating the role of rts1 in dna replication initiation |
| work_keys_str_mv | AT wallisa investigatingtheroleofrts1indnareplicationinitiation AT nieduszynskic investigatingtheroleofrts1indnareplicationinitiation |