{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,11,4]],"date-time":"2025-11-04T23:31:25Z","timestamp":1762299085443,"version":"3.40.5"},"reference-count":51,"publisher":"Wiley","issue":"2","license":[{"start":{"date-parts":[[2019,9,4]],"date-time":"2019-09-04T00:00:00Z","timestamp":1567555200000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"}],"funder":[{"DOI":"10.13039\/501100001824","name":"Grantov\u00e1 Agentura \u010cesk\u00e9 Republiky","doi-asserted-by":"publisher","award":["18\u201007748S"],"award-info":[{"award-number":["18\u201007748S"]}],"id":[{"id":"10.13039\/501100001824","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["febs.onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["The FEBS Journal"],"published-print":{"date-parts":[[2020,1]]},"abstract":"<jats:p><jats:italic>Streptococcus\u00a0pneumoniae<\/jats:italic>is a Gram\u2010positive bacterium that is a major agent of community\u2010acquired bacterial pneumonia, meningitis and sepsis. Although the mismatch repair function of<jats:italic>S.\u00a0pneumoniae<\/jats:italic>has been assigned to the<jats:italic>hexA\u2010hexB<\/jats:italic>gene products, an enzyme capable of the direct elimination of noncanonical nucleotides from the cytoplasm has not been described for this bacterium. Our results show that Spr1057, a protein with previously unknown function, is involved in the inactivation of mutagenic pyrimidine nucleotides and was accordingly designated PynA (pyrimidine nucleotidase A). Biochemical assays confirmed the phosphatase activity of the recombinant enzyme and revealed its metal ion dependence for optimal enzyme activity. We demonstrated that PynA forms a homodimer with higher<jats:italic>in\u00a0vitro<\/jats:italic>activity towards noncanonical 5\u2010fluoro\u20102\u2032\u2010deoxyuridine monophosphate than towards canonical thymidine monophosphate. Furthermore, we showed via<jats:italic>in\u00a0vivo<\/jats:italic>assays that PynA protects cells against noncanonical pyrimidine derivatives such as 5\u2010fluoro\u20102\u2032\u2010deoxyuridine and prevents the incorporation of the potentially mutagenic 5\u2010bromo\u20102\u2032\u2010deoxyuridine (5\u2010BrdU) into<jats:styled-content style=\"fixed-case\">DNA<\/jats:styled-content>. Fluctuation analysis performed under<jats:italic>S.\u00a0pneumoniae<\/jats:italic>exposure to 5\u2010BrdU revealed that the<jats:italic>pynA<\/jats:italic>null strain accumulates random mutations with high frequency, resulting in a 30\u2010fold increase in the mutation rate. The data support a model in which PynA, a protein conserved in other Gram\u2010positive bacteria, functions as a house\u2010cleaning enzyme by selectively eliminating noncanonical nucleotides and maintaining the purity of<jats:styled-content style=\"fixed-case\">dNTP<\/jats:styled-content>pools, similar to the YjjG protein described for<jats:italic>Escherichia\u00a0coli<\/jats:italic>.<\/jats:p>","DOI":"10.1111\/febs.15049","type":"journal-article","created":{"date-parts":[[2019,8,22]],"date-time":"2019-08-22T17:30:39Z","timestamp":1566495039000},"page":"267-283","update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":5,"title":["PynA is a pyrimidine 5\u2032\u2010nucleotidase that functions as an antimutator protein in<i>Streptococcus\u00a0pneumoniae<\/i>"],"prefix":"10.1111","volume":"287","author":[{"given":"Ale\u0161","family":"Ulrych","sequence":"first","affiliation":[{"name":"Institute of Microbiology, v.v.i. 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