{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2023,7,24]],"date-time":"2023-07-24T10:51:18Z","timestamp":1690195878261},"reference-count":20,"publisher":"Oxford University Press (OUP)","issue":"4","license":[{"start":{"date-parts":[[2022,7,5]],"date-time":"2022-07-05T00:00:00Z","timestamp":1656979200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/academic.oup.com\/journals\/pages\/open_access\/funder_policies\/chorus\/standard_publication_model"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"published-print":{"date-parts":[[2022,9,30]]},"abstract":"<jats:title>Abstract<\/jats:title>\n               <jats:p>Ribonuclease (RNase) H2 is involved in the removal of ribonucleotides embedded in genomic DNA. Eukaryotic RNase H2 is a heterotrimer consisting of the catalytic A subunit (RH2A) and the accessory B and C subunits. This study aimed to compare the cellular activities of wild-type ribonuclease (RNase) H2 and its variants with a mutation causing neuroinflammatory autoimmune disease, Aicardi\u2013Gouti\u00e8res syndrome (AGS). We first analyzed cellular RNase H2 activity and ribonucleotide content in the genomic DNA of RH2A-knockout (KO) mouse fibroblast NIH3T3 cells after transfection with a transient expression plasmid encoding mouse wild-type RH2A. From 4\u00a0h after transfection, the RNase H2 activity increased and the amount of ribonucleotides decreased, as compared with the corresponding non-transfected RH2A-KO cells. This demonstrated the rapidness of ribonucleotide turnover in mammalian genomic DNA and the importance of continuous expression of RNase H2 to maintain the ribonucleotide amount low. Next, we expressed mouse RH2A variants with a mutation corresponding to a human AGS-causing mutation in RH2A-KO NIH3T3 cells. Neither increase in RNase H2 activity nor decrease in ribonucleotide amount was observed for G37S; however, both conditions were observed for N213I and R293H. This corresponded with our previous results on the activity of recombinant human RNase H2 variants.<\/jats:p>","DOI":"10.1093\/jb\/mvac056","type":"journal-article","created":{"date-parts":[[2022,7,6]],"date-time":"2022-07-06T07:59:53Z","timestamp":1657094393000},"page":"225-231","source":"Crossref","is-referenced-by-count":1,"title":["Analysis of ribonucleotide content in the genomic DNA of ribonuclease H2 A subunit (RH2A)-knockout NIH3T3 cells after transient expression of wild-type RH2A or RH2A variants with an Aicardi\u2013Gouti\u00e8res syndrome-causing mutation"],"prefix":"10.1093","volume":"172","author":[{"given":"Mako","family":"Kandabashi","sequence":"first","affiliation":[{"name":"Graduate School of Agriculture Division of Food Science and Biotechnology, , Kyoto University, Sakyo-ku, Kyoto 606-8502, Japan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Haruna","family":"Yano","sequence":"additional","affiliation":[{"name":"Graduate School of Agriculture Division of Food Science and Biotechnology, , Kyoto University, Sakyo-ku, Kyoto 606-8502, Japan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Haruka","family":"Hara","sequence":"additional","affiliation":[{"name":"Graduate School of Agriculture Division of Food Science and Biotechnology, , Kyoto University, Sakyo-ku, Kyoto 606-8502, Japan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Saori","family":"Ogawa","sequence":"additional","affiliation":[{"name":"Graduate School of Agriculture Division of Food Science and Biotechnology, , Kyoto University, Sakyo-ku, Kyoto 606-8502, Japan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Kana","family":"Kamoda","sequence":"additional","affiliation":[{"name":"Graduate School of Agriculture Division of Food Science and Biotechnology, , Kyoto University, Sakyo-ku, Kyoto 606-8502, Japan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Shu","family":"Ishibashi","sequence":"additional","affiliation":[{"name":"Graduate School of Agriculture Division of Food Science and Biotechnology, , Kyoto University, Sakyo-ku, Kyoto 606-8502, Japan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Kohei","family":"Himeda","sequence":"additional","affiliation":[{"name":"Graduate School of Agriculture Division of Food Science and Biotechnology, , Kyoto University, Sakyo-ku, Kyoto 606-8502, Japan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Misato","family":"Baba","sequence":"additional","affiliation":[{"name":"Graduate School of Agriculture Division of Food Science and Biotechnology, , Kyoto University, Sakyo-ku, Kyoto 606-8502, Japan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Teisuke","family":"Takita","sequence":"additional","affiliation":[{"name":"Graduate School of Agriculture Division of Food Science and Biotechnology, , Kyoto University, Sakyo-ku, Kyoto 606-8502, Japan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Kiyoshi","family":"Yasukawa","sequence":"additional","affiliation":[{"name":"Graduate School of Agriculture Division of Food Science and Biotechnology, , Kyoto University, Sakyo-ku, Kyoto 606-8502, Japan"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"286","published-online":{"date-parts":[[2022,7,5]]},"reference":[{"key":"2022100315383759100_ref1","doi-asserted-by":"crossref","first-page":"1494","DOI":"10.1111\/j.1742-4658.2009.06908.x","article-title":"Ribonuclease H: the enzymes in eukaryotes","volume":"276","author":"Cerritelli","year":"2009","journal-title":"FEBS J."},{"key":"2022100315383759100_ref2","doi-asserted-by":"crossref","first-page":"1506","DOI":"10.1111\/j.1742-4658.2009.06909.x","article-title":"Ribonuclease H: properties, substrate specificity and roles in retroviral reverse transcription","volume":"276","author":"Champoux","year":"2009","journal-title":"FEBS J."},{"key":"2022100315383759100_ref3","doi-asserted-by":"crossref","first-page":"1482","DOI":"10.1111\/j.1742-4658.2009.06907.x","article-title":"Ribonuclease H: molecular diversities, substrate biding domains, and catalytic mechanism of the prokaryotic enzymes","volume":"276","author":"Tadokoro","year":"2009","journal-title":"FEBS J."},{"key":"2022100315383759100_ref4","doi-asserted-by":"crossref","first-page":"980","DOI":"10.1016\/j.molcel.2012.06.035","article-title":"RNase H2-initiated ribonucleotide excision repair","volume":"47","author":"Sparks","year":"2012","journal-title":"Mol. 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