Translation of human Δ133p53 mRNA and its targeting by antisense oligonucleotides complementary to the 5'-terminal region of this mRNA
| Autor: | Dorota Gurda-Woźna, Paulina Żydowicz-Machtel, Agata Swiatkowska, Jerzy Ciesiołka, Mariola Dutkiewicz |
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| Jazyk: | angličtina |
| Rok vydání: | 2021 |
| Předmět: |
Gene isoform
Small interfering RNA Cell Survival Science Materials Science Oligonucleotides Gene Expression Codon Initiator Transfection Research and Analysis Methods Biochemistry Eukaryotic translation Genetics Humans Protein Isoforms RNA Messenger Molecular Biology Techniques Materials Molecular Biology Antisense Oligonucleotides Gene Antisense RNA Translation Initiation Messenger RNA Multidisciplinary Biology and life sciences Nucleotides Chemistry Intron Translation (biology) Oligonucleotides Antisense HCT116 Cells Introns Cell biology Nucleic acids Oligomers Protein Biosynthesis Physical Sciences RNA Medicine Protein Translation Transcription Initiation Site Tumor Suppressor Protein p53 Research Article |
| Zdroj: | PLoS ONE, Vol 16, Iss 9, p e0256938 (2021) PLoS ONE |
| ISSN: | 1932-6203 |
| Popis: | The p53 protein is expressed as at least twelve protein isoforms. Within intron 4 of the human TP53 gene, a P2 transcription initiation site is located and this transcript encodes two p53 isoforms: Δ133p53 and Δ160p53. Here, the secondary structure of the 5′-terminal region of P2-initiated mRNA was characterized by means of the SHAPE and Pb2+-induced cleavage methods and for the first time, a secondary structure model of this region was proposed. Surprisingly, only Δ133p53 isoform was synthetized in vitro from the P2-initiated p53 mRNA while translation from both initiation codons occurred after the transfection of vector-encoded model mRNA to HCT116 cells. Interestingly, translation performed in the presence of the cap analogue suggested that the cap-independent process contributes to the translation of P2-initiated p53 mRNA. Subsequently, several antisense oligonucleotides targeting the 5′-terminal region of P2-initiated p53 mRNA were designed. The selected oligomers were applied in in vitro translation assays as well as in cell lines and their impact on the Δ133p53 synthesis and on cell viability was investigated. The results show that these oligomers are attractive tools in the modulation of the translation of P2-initiated p53 mRNA through attacking the 5′ terminus of the transcript. Since cell proliferation is also reduced by antisense oligomers that lower the level of Δ133p53, this demonstrates an involvement of this isoform in tumorigenesis. |
| Databáze: | OpenAIRE |
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