AP lyase activity of the human ribosomal protein uS3: The DNA cleavage sequence specificity and the location of the enzyme active center Научная публикация
Журнал |
Biochimica et Biophysica Acta - Proteins and Proteomics
ISSN: 1570-9639 |
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Вых. Данные | Год: 2023, Том: 1871, Номер: 2, Номер статьи : 140880, Страниц : DOI: 10.1016/j.bbapap.2022.140880 | ||
Ключевые слова | Human ribosomal protein uS3 Single-stranded DNA Apurin/apyrimidine site AP lyase activity Sequence specificity Cross-linking Active site | ||
Авторы |
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Организации |
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Реферат:
The human protein uS3, a component of the small ribosomal subunit, has a long-known extra-ribosomal activity
as an enzyme of base excision DNA repair displayed in its ability to cleave DNA at abasic (AP) sites. It has been
found that the efficacy of DNA cleavage by uS3 in vitro depends on the DNA sequence. To clarify the issue on the
sequence specificity of uS3 as an AP lyase in general, we applied a combinatorial approach based on the use of a
model single-stranded circular DNA with an AP site flanked with random trinucleotides at both sides. The
cleavage of this DNA by uS3 under conditions when only its minor portion undergoes the reaction resulted in the
formation of the linear DNA with random triplets at the 5′ and 3′ termini. NGS sequencing of the DNA library
derived from this DNA allowed identifying the contexts within which uS3 cleaves DNA the most and the least
effectively. Given that the AP lyase reaction occurs via the formation of a covalent intermediate (Schiff base), we
determined the region comprising the active center of the uS3 protein. By digesting of uS3 cross-linked to a
radiolabeled AP site-containing model DNA with specific proteolytic agents followed by analysis of the resulting
modified oligopeptides, the cross-link was mapped to the region 155–192 (likely, to R173/R178). Thus, our
results clarified two previously unstudied features of the uS3 AP lyase activity, one related to the recognition of
sequences in DNA surrounding the AP site, and the other to the protein region directly contacting this site.
Библиографическая ссылка:
Ochkasova A.
, Arbuzov G.D.
, Kabilov M.R.
, Tupikin A.E.
, Karpova G.G.
, Graifer D.M.
AP lyase activity of the human ribosomal protein uS3: The DNA cleavage sequence specificity and the location of the enzyme active center
Biochimica et Biophysica Acta - Proteins and Proteomics. 2023. V.1871. N2. 140880 . DOI: 10.1016/j.bbapap.2022.140880 WOS Scopus
AP lyase activity of the human ribosomal protein uS3: The DNA cleavage sequence specificity and the location of the enzyme active center
Biochimica et Biophysica Acta - Proteins and Proteomics. 2023. V.1871. N2. 140880 . DOI: 10.1016/j.bbapap.2022.140880 WOS Scopus
Даты:
Опубликована online: | 15 нояб. 2022 г. |
Идентификаторы БД:
Web of science | WOS:000890307400002 |
Scopus | 2-s2.0-85142208914 |
PMID (PubMed) | 36396097 |
OpenAlex | W4308951091 |
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