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- Title
Construction and characterization of ribonuclease H2 C subunit-knockout NIH3T3 cells.
- Authors
Haruka Hara; Haruna Yano; Kaho Akazawa; Kana Kamoda; Mako Kandabashi; Misato Baba; Teisuke Takita; Kiyoshi Yasukawa
- Abstract
Mammalian ribonuclease (RNase) H2 is a trimer consisting of catalytic A and accessory B and C subunits. RNase H2 is involved in the removal of misincorporated ribonucleotides from genomic DNA. In humans, mutations in RNase H2 gene cause a severe neuroinflammatory disorder, Aicardi–Goutières syndrome (AGS). Here, we constructed RNase H2 C subunit (RH2C)-knockout mouse fibroblast NIH3T3 cells. Compared with the wild-type NIH3T3 cells, the knockout cells exhibited a decreased single ribonucleotide-hydrolyzing activity and an increased accumulation of ribonucleotides in genomic DNA. Transient expression of wild-type RH2C in the knockout cells increased this activity and decreased this ribonucleotide accumulation. Same events were observed when RH2C variants with an AGS-causing mutation, R69W or K145I, were expressed. These results corresponded with our previous results on the RNase H2 A subunit (RH2A)-knockout NIH3T3 cells and the expression of wild-type RH2A or RH2A variants with an AGS-causing mutation, N213I and R293H, in the RH2A-knockout cells.
- Subjects
RIBONUCLEASES; NEUROLOGICAL disorders; RIBONUCLEOTIDES; DNA
- Publication
Bioscience, Biotechnology & Biochemistry, 2023, Vol 87, Issue 8, p865
- ISSN
0916-8451
- Publication type
Article
- DOI
10.1093/bbb/zbad077