Direct visualization of nucleolar G-quadruplexes in live cells by using a fluorescent light-up probe.
Biochim Biophys Acta Gen Subj
; 1862(5): 1101-1106, 2018 May.
Article
in En
| MEDLINE
| ID: mdl-29410183
ABSTRACT
BACKGROUND:
Direct detection of G-quadruplexes in human cells has become an important issue due to the vital role of G-quadruplex related to biological functions. Despite several probes have been developed for detection of the G-quadruplexes in cytoplasm or whole cells, the probe being used to monitor the nucleolar G-quadruplexes is still lacking.METHODS:
Formation of the nucleolar G-quadruplex structures was confirmed by using circular dichroism (CD) spectroscopy. The binding affinity and selectivity of Thioflavin T (ThT) towards various DNA/RNA motifs in solution and gel system were measured by using fluorescence spectroscopy and polyacrylamide gel electrophoresis (PAGE), respectively. G-quadruplex imaging in live cells was directly captured by using confocal laser scanning microscopy (CLSM).RESULTS:
Formation of the rDNA and rRNA G-quadruplex structures is demonstrated in vitro. ThT is found to show much higher affinity and selectivity towards these G-quadruplex structures versus other nucleic acid motifs either in solution or in gel system. The nucleolar G-quadruplexes in living cells are visualized by using ThT as a fluorescent probe. G-quadruplex-ligand treatments in live cells lead to sharp decrease of ThT signal.CONCLUSIONS:
The natural existence of the G-quadruplexes structure in the nucleoli of living cells is directly visualized by using ThT as an indicator. GENERALSIGNIFICANCE:
The research provides substantive evidence for formation of the rRNA G-quadruplex structures, and also offers an effective probe for direct visualization of the nucleolar G-quadruplexes in living cells.Key words
Full text:
1
Collection:
01-internacional
Database:
MEDLINE
Main subject:
Thiazoles
/
Molecular Probes
/
Cell Nucleus
/
G-Quadruplexes
/
Fluorescent Dyes
Limits:
Humans
Language:
En
Journal:
Biochim Biophys Acta Gen Subj
Year:
2018
Type:
Article