CiAPEX2 and CiP0, candidates of AP endonucleases in Ciona intestinalis, have 3′-5′ exonuclease activity and contribute to protection against oxidative stress
Abstract Apurinic/apyrimidinic (AP) sites are one of the most frequent DNA lesions. AP sites inhibit transcription and DNA replication, and induce cell death. AP endonucleases are key enzymes in AP site repair. Several types of AP endonucleases have been reported, such as AP endonuclease 2 (APEX2) a...
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| Format: | Article |
| Language: | English |
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BMC
2017-12-01
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| Series: | Genes and Environment |
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| Online Access: | http://link.springer.com/article/10.1186/s41021-017-0087-7 |
| _version_ | 1818539593302016000 |
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| author | Masafumi Funakoshi Daisuke Nambara Yuichiro Hayashi Qiu-Mei Zhang-Akiyama |
| author_facet | Masafumi Funakoshi Daisuke Nambara Yuichiro Hayashi Qiu-Mei Zhang-Akiyama |
| author_sort | Masafumi Funakoshi |
| collection | DOAJ |
| description | Abstract Apurinic/apyrimidinic (AP) sites are one of the most frequent DNA lesions. AP sites inhibit transcription and DNA replication, and induce cell death. AP endonucleases are key enzymes in AP site repair. Several types of AP endonucleases have been reported, such as AP endonuclease 2 (APEX2) and ribosomal protein P0 (P0). However, it is not known how the functions and roles differ among AP endonucleases. To clarify the difference of roles among AP endonucleases, we conducted biochemical analysis focused on APEX2 and P0 homologues in Ciona intestinalis. Amino acid sequence analysis suggested that CiAPEX2 and CiP0 are AP endonuclease homologues. Although we could not detect AP endonuclease or 3′-phosphodiesterase activity, these two purified proteins exhibited 3′-5′ exonuclease activity. This 3′-5′ exonuclease activity was sensitive to ethylenediaminetetraacetic acid (EDTA), and the efficiency of this activity was influenced by the 3′-terminus of substrate DNA. Both CiAPEX2 and CiP0 degraded not only a 5′-protruding DNA end, but also nicked DNA, which is generated through AP endonuclease 1 (APEX1) cleavage. These two genes partially complemented the growth rate of AP endonuclease-deficient Escherichia coli treated with hydrogen peroxide. These results indicate that 3′-5′ exonuclease activity is an evolutionarily conserved enzymatic activity of APEX2 and P0 homologues and this enzymatic activity may be important for AP endonucleases. |
| first_indexed | 2024-12-11T21:44:07Z |
| format | Article |
| id | doaj.art-be9332d85cc1484c9c1ec6a0ac0bf410 |
| institution | Directory Open Access Journal |
| issn | 1880-7062 |
| language | English |
| last_indexed | 2024-12-11T21:44:07Z |
| publishDate | 2017-12-01 |
| publisher | BMC |
| record_format | Article |
| series | Genes and Environment |
| spelling | doaj.art-be9332d85cc1484c9c1ec6a0ac0bf4102022-12-22T00:49:43ZengBMCGenes and Environment1880-70622017-12-013911810.1186/s41021-017-0087-7CiAPEX2 and CiP0, candidates of AP endonucleases in Ciona intestinalis, have 3′-5′ exonuclease activity and contribute to protection against oxidative stressMasafumi Funakoshi0Daisuke Nambara1Yuichiro Hayashi2Qiu-Mei Zhang-Akiyama3Laboratory of Stress Response Biology, Division of Biological Sciences, Graduate School of Science, Kyoto UniversityLaboratory of Stress Response Biology, Division of Biological Sciences, Graduate School of Science, Kyoto UniversityLaboratory of Stress Response Biology, Division of Biological Sciences, Graduate School of Science, Kyoto UniversityLaboratory of Stress Response Biology, Division of Biological Sciences, Graduate School of Science, Kyoto UniversityAbstract Apurinic/apyrimidinic (AP) sites are one of the most frequent DNA lesions. AP sites inhibit transcription and DNA replication, and induce cell death. AP endonucleases are key enzymes in AP site repair. Several types of AP endonucleases have been reported, such as AP endonuclease 2 (APEX2) and ribosomal protein P0 (P0). However, it is not known how the functions and roles differ among AP endonucleases. To clarify the difference of roles among AP endonucleases, we conducted biochemical analysis focused on APEX2 and P0 homologues in Ciona intestinalis. Amino acid sequence analysis suggested that CiAPEX2 and CiP0 are AP endonuclease homologues. Although we could not detect AP endonuclease or 3′-phosphodiesterase activity, these two purified proteins exhibited 3′-5′ exonuclease activity. This 3′-5′ exonuclease activity was sensitive to ethylenediaminetetraacetic acid (EDTA), and the efficiency of this activity was influenced by the 3′-terminus of substrate DNA. Both CiAPEX2 and CiP0 degraded not only a 5′-protruding DNA end, but also nicked DNA, which is generated through AP endonuclease 1 (APEX1) cleavage. These two genes partially complemented the growth rate of AP endonuclease-deficient Escherichia coli treated with hydrogen peroxide. These results indicate that 3′-5′ exonuclease activity is an evolutionarily conserved enzymatic activity of APEX2 and P0 homologues and this enzymatic activity may be important for AP endonucleases.http://link.springer.com/article/10.1186/s41021-017-0087-7AP endonucleaseRibosomal protein P03′-5′ exonuclease activityCiona intestinalis |
| spellingShingle | Masafumi Funakoshi Daisuke Nambara Yuichiro Hayashi Qiu-Mei Zhang-Akiyama CiAPEX2 and CiP0, candidates of AP endonucleases in Ciona intestinalis, have 3′-5′ exonuclease activity and contribute to protection against oxidative stress Genes and Environment AP endonuclease Ribosomal protein P0 3′-5′ exonuclease activity Ciona intestinalis |
| title | CiAPEX2 and CiP0, candidates of AP endonucleases in Ciona intestinalis, have 3′-5′ exonuclease activity and contribute to protection against oxidative stress |
| title_full | CiAPEX2 and CiP0, candidates of AP endonucleases in Ciona intestinalis, have 3′-5′ exonuclease activity and contribute to protection against oxidative stress |
| title_fullStr | CiAPEX2 and CiP0, candidates of AP endonucleases in Ciona intestinalis, have 3′-5′ exonuclease activity and contribute to protection against oxidative stress |
| title_full_unstemmed | CiAPEX2 and CiP0, candidates of AP endonucleases in Ciona intestinalis, have 3′-5′ exonuclease activity and contribute to protection against oxidative stress |
| title_short | CiAPEX2 and CiP0, candidates of AP endonucleases in Ciona intestinalis, have 3′-5′ exonuclease activity and contribute to protection against oxidative stress |
| title_sort | ciapex2 and cip0 candidates of ap endonucleases in ciona intestinalis have 3 5 exonuclease activity and contribute to protection against oxidative stress |
| topic | AP endonuclease Ribosomal protein P0 3′-5′ exonuclease activity Ciona intestinalis |
| url | http://link.springer.com/article/10.1186/s41021-017-0087-7 |
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