{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,4,4]],"date-time":"2026-04-04T12:49:01Z","timestamp":1775306941430,"version":"3.50.1"},"reference-count":98,"publisher":"Springer Science and Business Media LLC","issue":"1","content-domain":{"domain":["link.springer.com"],"crossmark-restriction":false},"short-container-title":["BMC Syst Biol"],"published-print":{"date-parts":[[2013,12]]},"abstract":"<jats:title>Abstract<\/jats:title>\n          <jats:sec>\n            <jats:title>Background<\/jats:title>\n            <jats:p>The genome of living organisms is constantly exposed to several damaging agents that induce different types of DNA lesions, leading to cellular malfunctioning and onset of many diseases. To maintain genome stability, cells developed various repair and tolerance systems to counteract the effects of DNA damage. Here we focus on Post Replication Repair (PRR), the pathway involved in the bypass of DNA lesions induced by sunlight exposure and UV radiation. PRR acts through two different mechanisms, activated by mono- and poly-ubiquitylation of the DNA sliding clamp, called Proliferating Cell Nuclear Antigen (PCNA).<\/jats:p>\n          <\/jats:sec>\n          <jats:sec>\n            <jats:title>Results<\/jats:title>\n            <jats:p>We developed a novel protocol to measure the time-course ratios between mono-, di- and tri-ubiquitylated PCNA isoforms on a single western blot, which were used as the wet readout for PRR events in wild type and mutant <jats:italic>S. cerevisiae<\/jats:italic> cells exposed to acute UV radiation doses. Stochastic simulations of PCNA ubiquitylation dynamics, performed by exploiting a novel mechanistic model of PRR, well fitted the experimental data at low UV doses, but evidenced divergent behaviors at high UV doses, thus driving the design of further experiments to verify new hypothesis on the functioning of PRR. The model predicted the existence of a UV dose threshold for the proper functioning of the PRR model, and highlighted an overlapping effect of Nucleotide Excision Repair (the pathway effectively responsible to clean the genome from UV lesions) on the dynamics of PCNA ubiquitylation in different phases of the cell cycle. In addition, we showed that ubiquitin concentration can affect the rate of PCNA ubiquitylation in PRR, offering a possible explanation to the DNA damage sensitivity of yeast strains lacking deubiquitylating enzymes.<\/jats:p>\n          <\/jats:sec>\n          <jats:sec>\n            <jats:title>Conclusions<\/jats:title>\n            <jats:p>We exploited an <jats:italic>in vivo<\/jats:italic> and <jats:italic>in silico<\/jats:italic> combinational approach to analyze for the first time in a Systems Biology context the events of PCNA ubiquitylation occurring in PRR in budding yeast cells. Our findings highlighted an intricate functional crosstalk between PRR and other events controlling genome stability, and evidenced that PRR is more complicated and still far less characterized than previously thought.<\/jats:p>\n          <\/jats:sec>","DOI":"10.1186\/1752-0509-7-24","type":"journal-article","created":{"date-parts":[[2013,3,20]],"date-time":"2013-03-20T19:14:21Z","timestamp":1363806861000},"update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":5,"title":["In vivo and in silico analysis of PCNA ubiquitylation in the activation of the Post Replication Repair pathway in S. cerevisiae"],"prefix":"10.1186","volume":"7","author":[{"given":"Flavio","family":"Amara","sequence":"first","affiliation":[]},{"given":"Riccardo","family":"Colombo","sequence":"additional","affiliation":[]},{"given":"Paolo","family":"Cazzaniga","sequence":"additional","affiliation":[]},{"given":"Dario","family":"Pescini","sequence":"additional","affiliation":[]},{"given":"Attila","family":"Csik\u00e1sz-Nagy","sequence":"additional","affiliation":[]},{"given":"Marco Muzi","family":"Falconi","sequence":"additional","affiliation":[]},{"given":"Daniela","family":"Besozzi","sequence":"additional","affiliation":[]},{"given":"Paolo","family":"Plevani","sequence":"additional","affiliation":[]}],"member":"297","published-online":{"date-parts":[[2013,3,20]]},"reference":[{"key":"1090_CR1","doi-asserted-by":"publisher","first-page":"366","DOI":"10.1038\/35077232","volume":"411","author":"J Hoeijmakers","year":"2001","unstructured":"Hoeijmakers J: Genome maintenance mechanisms for preventing cancer. 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