Effect of recombinant nucleophosmin 1 analogue on artificial RNA internalization into human adenocarcinoma MCF-7 cells

• Published in: Russian journal of bioorganic chemistry 2014, Vol.40 (1), p.48-55
• Main Authors: Savelyeva, A. V., Semenov, D. V., Stepanov, G. A., Baryakin, D. N., Kuligina, E. V., Rabinov, I. V., Koval, O. A., Richter, V. A.
• Format: Article
• Language: English
• Subjects: Biochemistry; Biomedical and Life Sciences; Biomedicine; Bioorganic Chemistry; Life Sciences; Organic Chemistry
• ISSN: 1068-1620; 1608-330X
• DOI: 10.1134/S1068162014010099

In this study we obtained and characterized the recombinant analogue (Npm1-His 6 ) of multifunctional nucleolar phosphoprotein nucleophosmin 1 (NPM1) involved in crucial cellular processes such as transcription, reparation, and mitosis. The interaction of Npm1-His 6 with artificial noncoding RNAs in vitro and its influence on extracellular RNA accumulation in human adenocarcinoma MCF-7 cells were analyzed. It was found that the incubation of Npm1-His 6 with AluY RNA ( n > 300 nt) or U24 snoRNA analogues ( n ∼ 80 nt) but not with a short oligoribonucleotide ( n = 22 nt) resulted in the formation of the RNA-protein non-covalent complexes. Recombinant protein Npm1-His 6 was shown to increase the transfection efficacy of RNA with a complex secondary structure into MCF-7 human cells. The results allow us to conclude that nucleophosmin 1 not only binds RNAs with complex secondary structure but also promotes the uptake and internalization of these RNAs by human cells.