Wani, S and Maharshi, N and Kothiwal, D and Mahendrawada, L and Kalaivani, R and Laloraya, S (2018) Interaction of the Saccharomyces cerevisiae RING-domain protein Nse1 with Nse3 and the Smc5/6 complex is required for chromosome replication and stability. In: Current Genetics, 64 (3). pp. 599-617.
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Abstract
Genomic stability is maintained by the concerted actions of numerous protein complexes that participate in chromosomal duplication, repair, and segregation. The Smc5/6 complex is an essential multi-subunit complex crucial for repair of DNA double-strand breaks. Two of its subunits, Nse1 and Nse3, are homologous to the RING-MAGE complexes recently described in human cells. We investigated the contribution of the budding yeast Nse1 RING-domain by isolating a mutant nse1-103 bearing substitutions in conserved Zinc-coordinating residues of the RING-domain that is hypersensitive to genotoxic stress and temperature. The nse1-103 mutant protein was defective in interaction with Nse3 and other Smc5/6 complex subunits, Nse4 and Smc5. Chromosome loss was enhanced, accompanied by a delay in the completion of replication and a modest defect in sister chromatid cohesion, in nse1-103. The nse1-103 mutant was synthetic sick with rrm3∆ (defective in fork passage through pause sites), this defect was rescued by inactivation of Tof1, a subunit of the fork protection complex that enforces pausing. The temperature sensitivity of nse1-103 was partially suppressed by deletion of MPH1, encoding a DNA-helicase. Homology modeling of the structure of the budding yeast Nse1–Nse3 heterodimer based on the human Nse1–MAGEG1 structure suggests a similar organization and indicates that perturbation of the Zn-coordinating cluster has the potential to allosterically alter structural elements at the Nse1/Nse3 interaction interface that may abrogate their association. Our findings demonstrate that the budding yeast Nse1 RING-domain organization is important for interaction with Nse3, which is crucial for completion of chromosomal replication, cohesion, and maintenance of chromosome stability.
Item Type: | Journal Article |
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Publication: | Current Genetics |
Publisher: | Springer Verlag |
Additional Information: | The copyright for this article belongs to the Springer Verlag. |
Keywords: | heterodimer; histidine; hydroxyurea; mesylic acid; nse1 protein; nse3 protein; RING finger protein; Saccharomyces cerevisiae protein; smc5 protein; smc6 protein; unclassified drug; fungal DNA; mutagenic agent; protein binding; Saccharomyces cerevisiae protein, amino acid sequence; Article; cell lysate; cell mutant; chromosomal instability; chromosome aberration; chromosome loss; chromosome replication; comparative study; controlled study; DNA binding; DNA content; DNA replication; fungus growth; gene interaction; high temperature; immunoprecipitation; nonhuman; polyacrylamide gel electrophoresis; priority journal; pulsed field gel electrophoresis; RING finger motif; Saccharomyces cerevisiae; sister chromatid; steady state; temperature sensitivity; ultraviolet radiation; Western blotting; yeast cell; biosynthesis; chromosome; genetics; human; metabolism; protein domain; Saccharomyces cerevisiae; temperature, Chromosomal Instability; Chromosomes, Fungal; DNA Replication; DNA, Fungal; Electrophoresis, Gel, Pulsed-Field; Humans; Mutagens; Protein Binding; Protein Domains; Saccharomyces cerevisiae; Saccharomyces cerevisiae Proteins; Temperature |
Department/Centre: | Division of Biological Sciences > Biochemistry Division of Biological Sciences > Molecular Biophysics Unit |
Date Deposited: | 11 Aug 2022 09:57 |
Last Modified: | 11 Aug 2022 09:57 |
URI: | https://eprints.iisc.ac.in/id/eprint/75539 |
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