Dataset Information

A Spotlight on Rad52 in Cyanidiophytina (Rhodophyta): A Relic in Algal Heritage.

ABSTRACT: The RADiation sensitive52 (RAD52) protein catalyzes the pairing between two homologous DNA sequences' double-strand break repair and meiotic recombination, mediating RAD51 loading onto single-stranded DNA ends, and initiating homologous recombination and catalyzing DNA annealing. This article reports the characterization of RAD52 homologs in the thermo-acidophilic Cyanidiophyceae whose genomes have undergone extensive sequencing. Database mining, phylogenetic inference, prediction of protein structure and evaluation of gene expression were performed in order to determine the functionality of the RAD52 protein in Cyanidiophyceae. Its current function in Cyanidiophytina could be related to stress damage response for thriving in hot and acidic environments as well as to the genetic variability of these algae, in which, conversely to extant Rhodophyta, sexual mating was never observed.

SUBMITTER: Del Mondo A

PROVIDER: S-EPMC6410040 | biostudies-literature | 2019 Feb

REPOSITORIES: biostudies-literature

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A Spotlight on Rad52 in Cyanidiophytina (Rhodophyta): A Relic in Algal Heritage.

Del Mondo Angelo A Iovinella Manuela M Petriccione Milena M Nunziata Angelina A Davis Seth J SJ Cioppa Diana D Ciniglia Claudia C

Plants (Basel, Switzerland) 20190219 2

The RADiation sensitive52 (RAD52) protein catalyzes the pairing between two homologous DNA sequences' double-strand break repair and meiotic recombination, mediating RAD51 loading onto single-stranded DNA ends, and initiating homologous recombination and catalyzing DNA annealing. This article reports the characterization of RAD52 homologs in the thermo-acidophilic Cyanidiophyceae whose genomes have undergone extensive sequencing. Database mining, phylogenetic inference, prediction of protein str ...[more]

PMID: 30791384

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Project description:DNA double-strand breaks (DSBs) are toxic forms of DNA damage that must be repaired to maintain genome integrity. Telomerase can act upon a DSB to create a de novo telomere, a process that interferes with normal repair and creates terminal deletions. We previously identified sequences in Saccharomyces cerevisiae (SiRTAs; Sites of Repair-associated Telomere Addition) that undergo unusually high frequencies of de novo telomere addition, even when the original chromosome break is several kilobases distal to the eventual site of telomerase action. Association of the single-stranded telomere binding protein Cdc13 with a SiRTA is required to stimulate de novo telomere addition. Because extensive resection must occur prior to Cdc13 binding, we utilized these sites to monitor the effect of proteins involved in homologous recombination. We find that telomere addition is significantly reduced in the absence of the Rad51 recombinase, while loss of Rad52, required for Rad51 nucleoprotein filament formation, has no effect. Deletion of RAD52 suppresses the defect of the rad51Δ strain, suggesting that Rad52 inhibits de novo telomere addition in the absence of Rad51. The ability of Rad51 to counteract this effect of Rad52 does not require DNA binding by Rad51, but does require interaction between the two proteins, while the inhibitory effect of Rad52 depends on its interaction with Replication Protein A (RPA). Intriguingly, the genetic interactions we report between RAD51 and RAD52 are similar to those previously observed in the context of checkpoint adaptation. Forced recruitment of Cdc13 fully restores telomere addition in the absence of Rad51, suggesting that Rad52, through its interaction with RPA-coated single-stranded DNA, inhibits the ability of Cdc13 to bind and stimulate telomere addition. Loss of the Rad51-Rad52 interaction also stimulates a subset of Rad52-dependent microhomology-mediated repair (MHMR) events, consistent with the known ability of Rad51 to prevent single-strand annealing.

Project description:Extracellular or "relic" DNA is one of the largest pools of nucleic acids in the biosphere. Relic DNA can influence a number of important ecological and evolutionary processes, but it may also affect estimates of microbial abundance and diversity, which has implications for understanding environmental, engineered, and host-associated ecosystems. We developed models capturing the fundamental processes that regulate the size and composition of the relic DNA pools to identify scenarios leading to biased estimates of biodiversity. Our models predict that bias increases with relic DNA pool size, but only when the species abundance distributions (SADs) of relic and intact DNA are distinct from one another. We evaluated our model predictions by quantifying relic DNA and assessing its contribution to bacterial diversity using 16S rRNA gene sequences collected from different ecosystem types, including soil, sediment, water, and the mammalian gut. On average, relic DNA made up 33% of the total bacterial DNA pool but exceeded 80% in some samples. Despite its abundance, relic DNA had a minimal effect on estimates of taxonomic and phylogenetic diversity, even in ecosystems where processes such as the physical protection of relic DNA are common and predicted by our models to generate bias. Our findings are consistent with the expectation that relic DNA from different taxa degrades at a constant and equal rate, suggesting that it may not fundamentally alter estimates of microbial diversity.IMPORTANCE The ability to rapidly obtain millions of gene sequences and transcripts from a range of environments has greatly advanced understanding of the processes that regulate microbial communities. However, nucleic acids extracted from complex samples do not come only from viable microorganisms. Dead microorganisms can generate large pools of relic DNA that distort insight into the ecology and evolution of microbial systems. Here, we develop a conceptual and quantitative framework for understanding how relic DNA influences the structure of microbiomes. Our theoretical models and empirical results demonstrate that a large relic DNA pool does not automatically lead to biased estimates of microbial diversity. Rather, relic DNA effects emerge in combination with microscale processes that alter the commonness and rarity of sequences found in heterogeneous DNA pools.

Dataset Information

A Spotlight on Rad52 in Cyanidiophytina (Rhodophyta): A Relic in Algal Heritage.

Publications

A Spotlight on Rad52 in Cyanidiophytina (Rhodophyta): A Relic in Algal Heritage.

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