Caspase-2 (Casp-2) is an enzyme that regulates the development of apoptosis upon alternative splicing of its mRNA. The long form of Casp-2 (Casp-2L) promotes apoptosis while the short form (Casp-2S) has decreased enzymatic activity and inhibits the development of apoptotic processes. However, very little is known about the mechanism of Casp-2 alternative splicing. Several endonucleases are known to participate in this process. The aim of this study was to determine the role of EndoG in regulation of Casp-2 alternative splicing. Strong correlation between expression levels of EndoG and Casp-2 splice-variants was found in CD4⁺ and CD8⁺ human T lymphocytes. Such correlation increased after incubation of these cells with etoposide. Increased expression of Casp-2S was determined during EndoG over-expression in CD4⁺ T-cells, after EndoG treatment of cell cytoplasm and nuclei and after nuclei incubation with EndoG digested cell RNA. Casp-2 alternative splicing was induced by a 60-mer RNA oligonucleotide in naked nuclei and in cells after transfection. The identified long non-coding RNA of 1016 nucleotides is the precursor of the 60-mer RNA oligonucleotide. Based on the results the following mechanism has been proposed. Casp-2 pre-mRNA is transcribed from the coding DNA strand while long non-coding RNA is transcribed from the template strand of the Casp-2 gene. EndoG digests long non-coding RNA and produces the 60-mer RNA oligonucleotide complementary to the Casp-2 pre-mRNA exon 9 and intron 9 junction place. Interaction of the 60-mer RNA oligonucleotide and Casp-2 pre-mRNA causes alternative splicing.
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Keywords: EndoG, alternative splicing, caspase-2, T lymphocytes
Citation:
Zhdanov D.D., Gladilina Yu.A., Shisparenok A.N. (2024) Apoptotic endonuclease EndoG induces alternative splicing of Caspase-2. Biomeditsinskaya Khimiya, 70(4), 218-230.
Zhdanov D.D. et al. Apoptotic endonuclease EndoG induces alternative splicing of Caspase-2 // Biomeditsinskaya Khimiya. - 2024. - V. 70. -N 4. - P. 218-230.
Zhdanov D.D. et al., "Apoptotic endonuclease EndoG induces alternative splicing of Caspase-2." Biomeditsinskaya Khimiya 70.4 (2024): 218-230.
Zhdanov, D. D., Gladilina, Yu. A., Shisparenok, A. N. (2024). Apoptotic endonuclease EndoG induces alternative splicing of Caspase-2. Biomeditsinskaya Khimiya, 70(4), 218-230.
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