Dataset Information

Post-translational Regulation of DNA Polymerase ?, a Connection to Damage-Induced Cohesion in Saccharomyces cerevisiae.

ABSTRACT: Double-strand breaks that are induced postreplication trigger establishment of damage-induced cohesion in Saccharomyces cerevisiae, locally at the break site and genome-wide on undamaged chromosomes. The translesion synthesis polymerase, polymerase ?, is required for generation of damage-induced cohesion genome-wide. However, its precise role and regulation in this process is unclear. Here, we investigated the possibility that the cyclin-dependent kinase Cdc28 and the acetyltransferase Eco1 modulate polymerase ? activity. Through in vitro phosphorylation and structure modeling, we showed that polymerase ? is an attractive substrate for Cdc28 Mutation of the putative Cdc28-phosphorylation site Ser14 to Ala not only affected polymerase ? protein level, but also prevented generation of damage-induced cohesion in vivo We also demonstrated that Eco1 acetylated polymerase ? in vitro Certain nonacetylatable polymerase ? mutants showed reduced protein level, deficient nuclear accumulation, and increased ultraviolet irradiation sensitivity. In addition, we found that both Eco1 and subunits of the cohesin network are required for cell survival after ultraviolet irradiation. Our findings support functionally important Cdc28-mediated phosphorylation, as well as post-translational modifications of multiple lysine residues that modulate polymerase ? activity, and provide new insights into understanding the regulation of polymerase ? for damage-induced cohesion.

SUBMITTER: Wu PS

PROVIDER: S-EPMC7768261 | biostudies-literature | 2020 Dec

REPOSITORIES: biostudies-literature

ACCESS DATA

Publications

Post-translational Regulation of DNA Polymerase η, a Connection to Damage-Induced Cohesion in <i>Saccharomyces cerevisiae</i>.

Wu Pei-Shang PS Enervald Elin E Joelsson Angelica A Palmberg Carina C Rutishauser Dorothea D Hällberg B Martin BM Ström Lena L

Genetics 20201008 4

Double-strand breaks that are induced postreplication trigger establishment of damage-induced cohesion in <i>Saccharomyces cerevisiae</i>, locally at the break site and genome-wide on undamaged chromosomes. The translesion synthesis polymerase, polymerase η, is required for generation of damage-induced cohesion genome-wide. However, its precise role and regulation in this process is unclear. Here, we investigated the possibility that the cyclin-dependent kinase Cdc28 and the acetyltransferase Ec ...[more]

PMID: 33033113

Similar Datasets

Project description:Saccharomyces cerevisiae myosin type II-deficient (myo1?) strains remain viable and divide, despite the absence of a cytokinetic ring, by activation of the PKC1-dependent cell wall integrity pathway (CWIP). Since the myo1? transcriptional fingerprint is a subset of the CWIP fingerprint, the myo1? strain may provide a simplified paradigm for cell wall stress survival.To explore the post-transcriptional regulation of the myo1? stress response, 1,301 differentially regulated ribosome-bound mRNAs were identified by microarray analysis of which 204 were co-regulated by transcription and translation. Four categories of mRNA were significantly affected - protein biosynthesis, metabolism, carbohydrate metabolism, and unknown functions. Nine genes of the 20 CWIP fingerprint genes were post-transcriptionally regulated. Down and up regulation of selected ribosomal protein and cell wall biosynthesis mRNAs was validated by their distribution in polysomes from wild type and myo1? strains. Western blot analysis revealed accumulation of the phosphorylated form of eukaryotic translation initiation factor 2 (eIF2?-P) and a reduction in the steady state levels of the translation initiation factor eIF4Gp in myo1? strains. Deletion of GCN2 in myo1? abolished eIF2?p phosphorylation, and showed a severe growth defect. The presence of P-bodies in myo1? strains suggests that the process of mRNA sequestration is active, however, the three representative down regulated RP mRNAs, RPS8A, RPL3 and RPL7B were present at equivalent levels in Dcp2p-mCh-positive immunoprecipitated fractions from myo1? and wild type cells. These same RP mRNAs were also selectively co-precipitated with eIF2?-P in myo1? strains.Quantitative analysis of ribosome-associated mRNAs and their polyribosome distributions suggests selective regulation of mRNA translation efficiency in myo1? strains. Inhibition of translation initiation factor eIF2? (eIF2?-P) in these strains was by Gcn2p-dependent phosphorylation. The increase in the levels of eIF2?-P; the genetic interaction between GCN2 and MYO1; and the reduced levels of eIF4Gp suggest that other signaling pathways, in addition to the CWIP, may be important for myo1? strain survival. Selective co-immunoprecipitation of RP mRNAs with eIF2?-P in myo1? strains suggests a novel mode of translational regulation. These results indicate that post-transcriptional control is important in the myo1? stress response and possibly other stresses in yeast.

Dataset Information

Post-translational Regulation of DNA Polymerase ?, a Connection to Damage-Induced Cohesion in Saccharomyces cerevisiae.

Publications

Post-translational Regulation of DNA Polymerase η, a Connection to Damage-Induced Cohesion in <i>Saccharomyces cerevisiae</i>.

Similar Datasets

OmicsDI is part of the ELIXIR infrastructure

Tweets