Time-resolved fluorescence of DAPI in solution and bound to polydeoxynucleotides

Barcellona, Maria Luisa; Cardiel, G.; Gratton, Enrico

doi:10.1016/0006-291x(90)91270-3

Cited by 83 publications

(73 citation statements)

References 20 publications

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“…M2915-6A transformed with pCXJ15-COX7 6A-͓pCXJ15-COX8͔ M2915-6A transformed with pCXJ15-COX8 6A-͓pCXJ15-COX9͔ M2915-6A transformed with pCXJ15-COX9 6A-͓pRS416-COX9͔ M2915-6A transformed with pRS416-COX9 BY4741/⌬cox9 b MATa,his3,leu2,met15,ura3,cox9⌬::KanMX4 6A/⌬cox9 M2915-6A with cox9⌬::KanMX4 ⌬cox9-͓pRS416-COX9͔ 6A/⌬cox9 transformed with pRS416-COX9 ⌬cox9-͓pCXJ15-COX9͔ 6A/⌬cox9 transformed with pCXJ15-COX9 (Pringle et al, 1989;Barcellona et al, 1990). In brief, yeast cells were harvested from the cultures, washed with water, and incubated for 20 min with DAPI, DiOC 6 , and PI at concentrations of 2 g/ml, 0.64 ng/ml, and 10 g/ml, respectively.…”

Section: M2915-6amentioning

confidence: 99%

Miltefosine Induces Apoptosis-Like Cell Death in Yeast via Cox9p in CytochromecOxidase

Zuo¹,

Djordjevic²,

Oei³

et al. 2011

Mol Pharmacol

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Miltefosine has antifungal properties and potential for development as a therapeutic for invasive fungal infections. However, its mode of action in fungi is poorly understood. We demonstrate that miltefosine is rapidly incorporated into yeast, where it penetrates the mitochondrial inner membrane, disrupting mitochondrial membrane potential and leading to an apoptosislike cell death. COX9, which encodes subunit VIIa of the cytochrome c oxidase (COX) complex in the electron transport chain of the mitochondrial membrane, was identified as a potential target of miltefosine from a genomic library screen of the model yeast Saccharomyces cerevisiae. When overexpressed in S. cerevisiae, COX9, but not COX7 or COX8, led to a miltefosine-resistant phenotype. The effect of miltefosine on COX activity was assessed in cells expressing different levels of COX9. Miltefosine inhibited COX activity in a dose-dependent manner in Cox9p-positive cells. This inhibition most likely contributed to the miltefosine-induced apoptosis-like cell death.

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Section: M2915-6amentioning

confidence: 99%

Miltefosine Induces Apoptosis-Like Cell Death in Yeast via Cox9p in CytochromecOxidase

Zuo¹,

Djordjevic²,

Oei³

et al. 2011

Mol Pharmacol

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“…Therefore, the classical blue DNA dye DAPI, which belongs to the indol family, is the dye of choice for staining the chromosomes in the nucleus of fixed and living cells. It could be shown that the binding of DAPI to DNA leads to a 20-fold increase of the fluorescence (11). However, since DAPI can only be excited with ultraviolet (UV) light, exposition of this fluorescent dye interferes with the viability of the probed cells and somewhat limits the accessible observation time.…”

Section: Introductionmentioning

confidence: 99%

Nanomechanics of Fluorescent DNA Dyes on DNA Investigated by Magnetic Tweezers

Wang

Sischka

Walhorn

et al. 2016

Biophysical Journal

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Fluorescent DNA dyes are broadly used in many biotechnological applications for detecting and imaging DNA in cells and gels. Their binding alters the structural and nanomechanical properties of DNA and affects the biological processes that are associated with it. Although interaction modes like intercalation and minor groove binding already have been identified, associated mechanic effects like local elongation, unwinding, and softening of the DNA often remain in question. We used magnetic tweezers to quantitatively investigate the impact of three DNA-binding dyes (YOYO-1, DAPI, and DRAQ5) in a concentration-dependent manner. By extending and overwinding individual, torsionally constrained, nickfree dsDNA molecules, we measured the contour lengths and molecular forces that allow estimation of thermodynamic and nanomechanical binding parameters. Whereas for YOYO-1 and DAPI the binding mechanisms could be assigned to bis-intercalation and minor groove binding, respectively, DRAQ5 exhibited both binding modes in a concentration-dependent manner.

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“…9.1i). Additionally, when bound to DNA, fluorescence emitted from DAPI is 20 times greater than that of unbound, greatly reducing background signals [23]. Thus, its selectivity for DNA and high cell permeability allow for efficient staining of nuclei with limited cytosolic fluorescence.…”

Section: ′6-diamidino-2-phenylindole (Dapi)mentioning

confidence: 99%