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Varizhuk, A. M.

Publications and source records attributed to Varizhuk, A. M..

2 recordsLinked to original sources

Alteration in the intracellular Na+/K+ ratio regulates gene expression through the shift of G-quadruplex dynamics in living cells

Alteration in the intracellular Na+i/K+i ratio can play an important regulatory role in many mammalian cells. Here, we demonstrated that the expression of some early response genes does depend on the intracellular Na+i/K+i ratio and that the inhibition of DNA-binding activity of transcription factors c-Fos and c-Jun attenuates the upregulation of Zfp36 and Egr1 genes in HeLa cells. Both replacement of intracellular Na+ and K+ by Li+ and the dissipation of Na+i/K+i gradient demonstrated a similar effect on the transcription of Fos, Jun, Zfp36, and Ptgs2. Oligonucleotide sequences from the Fos, Kit, and My[c] promoters with an ability to fold into G-quadruplexes (G4s) change their conformations differently depending on the ratio of Na+, K+, and Li+ ions in vitro. Fluorescence lifetime imaging microscopy studies using DNA G4-specific dye DAOTA-M2 revealed that changes in compositions and ratios of monovalent metal cations in HeLa cells were accompanied by alterations in intranuclear G4s dynamics. The data demonstrate that these structures may be considered as intracellular sensors of monovalent metal cations regulating gene expression through the shift of G4s dynamics.

biochemistry↗

Red light-emitting short Mango-based system enables tracking a mycobacterial small noncoding RNA in infected macrophages

Progress in the studies of RNA metabolism and function relies largely on molecular imaging systems, including those comprising a fluorogenic dye and an aptamer-based fluorescence-activating tag. G4 aptamers of Mango family, typically combined with a duplex/hairpin scaffold, activate fluorescence of a green-emitting dye TO1-biotin and hold great promise for intracellular RNA tracking. Here, we report a new Mango-based imaging platform. Its key advantages are tunability of spectral properties and applicability for visualization of small RNA molecules that require minimal tag size. The former advantage is due to an expanded (green-to-red-emitting) palette of TO1-inspired fluorogenic dyes, and the latter is ensured by the truncated duplex scaffold. To illustrate applicability of the improved platform, we tagged Mycobacterium tuberculosis sncRNA with the shortened aptamer-scaffold tag and visualized it in bacteria and bacteria-infected macrophages using the new red light-emitting Mango-activated dye.

synthetic biology↗