Differential requirement for the N-terminal catalytic domain of the DNA polymerase ε p255 subunit in the mitotic cell cycle and the endocycle
| Autor: | Kengo Sakaguchi, Mieko Kawai, Osamu Suyari, Masamitsu Yamaguchi, Hideki Yoshida, Hiroyuki Ida |
|---|---|
| Rok vydání: | 2011 |
| Předmět: |
DNA Replication
DNA polymerase DNA polymerase II Cell Cycle Proteins Gene Expression Regulation Enzymologic Salivary Glands S Phase Animals Genetically Modified chemistry.chemical_compound Mitotic cell cycle Catalytic Domain Genetics Endoreduplication Animals Drosophila Proteins RNA Double-Stranded DNA clamp biology Minichromosome Maintenance Proteins Cell Cycle DNA replication Gene Expression Regulation Developmental General Medicine DNA Polymerase II Molecular biology Protein Subunits chemistry Gene Knockdown Techniques Larva biology.protein Drosophila Primase DNA |
| Zdroj: | Gene. 495(2) |
| ISSN: | 1879-0038 |
| Popis: | In Drosophila, the 255 kDa catalytic subunit (dpolep255) and the 58 kDa subunit of DNA polymerase e (dpolep58) have been identified. The N-terminus of dpolep255 carries well-conserved six DNA polymerase subdomains and five 3′→5′ exonuclease motifs as observed with Pole in other species. We here examined roles of dpolep255 during Drosophila development using transgenic fly lines expressing double stranded RNA (dsRNA). Expression of dpolep255 dsRNA in eye discs induced a small eye phenotype and inhibited DNA synthesis, indicating a role in the G1–S transition and/or S-phase progression of the mitotic cycle. Similarly, expression of dpolep255 dsRNA in the salivary glands resulted in small size and endoreplication defects, demonstrating a critical role in endocycle progression. In the eye disc, defects induced by knockdown of dpolep255 were rescued by overexpression of the C-terminal region of dpolep255, indicating that the function of this non-catalytic domain is conserved between yeast and Drosophila. However, this was not the case for the salivary gland, suggesting that the catalytic N-terminal region is crucial for endoreplication and its defect cannot be complemented by other DNA polymerases. In addition, several genetic interactants with dpolep255 including genes related to DNA replication such as RFC, DNA primase, DNA polη, Mcm10 and Psf2 and chromatin remodeling such as Iswi were also identified. |
| Databáze: | OpenAIRE |
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