Effects of increased cellular zinc levels on gene and protein expression in HT-29 cells

Kindermann, Birgit; Döring, Frank; Fuchs, D.; Pfaffl, Michael W.; Daniel, Hannelore

doi:10.1007/s10534-005-1247-y

Cited by 23 publications

(9 citation statements)

References 49 publications

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“…Previous studies suggested that cellular targets of elevated intracellular free Zn are diverse and may include organelles such as mitochondria or lysosomes, particular enzymatic systems or individual nuclear genes (Dineley et al 2003;Kindermann et al 2005;Ostrakhovitch and Cherian 2005;Yu et al 2009). Moreover, available evidence in the literature suggests differing dynamics of this process in various models, with relative scarcity of data in solid tumor cells (Ostrakhovitch and Cherian 2004).…”

Section: Discussionmentioning

confidence: 99%

Zinc pyrithione induces cellular stress signaling and apoptosis in Hep-2 cervical tumor cells: the role of mitochondria and lysosomes

Rudolf

Červinka

2010

Biometals

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Increased intracellular free zinc concentrations are associated with activation of several stress signaling pathways, specific organelle injury and final cell death. In the present work we examined the involvement of mitochondria and lysosomes and their crosstalk in free zinc-induced cell demise. We report that treatment of cervical tumor Hep-2 cells with zinc pyrithione leads to an early appearance of cytoplasmic zinc-specific foci with corresponding accumulation of zinc first in mitochondria and later in lysosomes. Concomitant with these changes, upregulation of expression of metallothionein II A gene as well as the increased abundance of its protein occurs. Moreover, zinc activates p53 and its dependent genes including Puma and Bax and they contribute to an observed loss of mitochondrial membrane potential and activation of apoptosis. Conversely, lysosomal membrane permeabilization and its promoted cleavage of Bid occurs in a delayed manner in treated cells and their effect on decrease of mitochondrial membrane potential is limited. The use of specific inhibitors as well as siRNA technology suggest a crucial role of MT-IIA in trafficking of free zinc into mitochondria or lysosomes and regulation of apoptotic or necrotic cell demise.

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Section: Discussionmentioning

confidence: 99%

Zinc pyrithione induces cellular stress signaling and apoptosis in Hep-2 cervical tumor cells: the role of mitochondria and lysosomes

Rudolf

Červinka

2010

Biometals

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“…Application of this Zn 2+ chelating agent has been employed to induce functional zinc deficiency and demonstrate the role of intracellular Zn 2+ in various processes such as apoptosis, oxidant stress, properties of neuronal synaptic transmission, regulation of transcription factors such as NF-kappa β, etc. [47][48][49][50][51]. The assumption seems to be that TPEN sequesters "chelatable" or "accessible" Zn 2+ , defined vaguely as free or loosely bound Zn 2+ ; but the actual impact of TPEN on Zn-proteome has not previously been measured.…”

Section: Discussionmentioning

confidence: 99%

Zinc binding ligands and cellular zinc trafficking: Apo-metallothionein, glutathione, TPEN, proteomic zinc, and Zn-Sp1

Rana¹,

Kothinti²,

Meeusen³

et al. 2008

Journal of Inorganic Biochemistry

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Many cell types contain metal-ion unsaturated metallothionein (MT). Considering the Zn 2+ binding affinity of metallothionein, the existence of this species in the intracellular environment constitutes a substantial "thermodynamic sink." Indeed, the mM concentration of glutathione may be thought of in the same way. In order to understand how apo-MT and the rest of the Zn-proteome manage to co-exist, experiments examined the in vitro reactivity of Zn-proteome with apo-MT, glutathione (GSH), and a series of common Zn 2+ chelating agents including N,N,N',N'-(2-pyridylethyl) ethylenediammine (TPEN), EDTA, and [(2,2'-oxyproplylene-dinitrilo]tetraacetic acid (EGTA). Less than 10% of Zn-proteome from U87 mg cells reacted with apo-MT or GSH. In contrast, each of the synthetic chelators was 2−3 times more reactive. TPEN, a cell permeant reagent, also reacted rapidly with both Zn-proteome and Zn-MT in LLC-PK 1 cells. Taking a specific zinc finger protein for further study, apo-MT, GSH, and TPEN inhibited the binding of Zn 3 -Sp1 with its cognate DNA site (GC-1) in the sodium-glucose co-transporter promoter of mouse kidney. In contrast, preformation of Zn 3 -Sp1-(GC-1) prevented reaction with apo-MT and GSH; TPEN remained active but at a higher concentration. Whereas, Zn 3 -Sp1 is active in cells containing apo-MT and GSH, exposure of LLC-PK 1 cells to TPEN for 24 h largely inactivated its DNA binding activity. The results help to rationalize the steady state presence of cellular apo-MT in the midst of the many, diverse members of the Znproteome. They also show that TPEN is a robust intracellular chelator of proteomic Zn 2+ .

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“…For example, zinc(II) is essential for many cellular processes, including DNA synthesis, transcription, and translation, but its excess can be toxic (Babich and Stotzky 1978;Kindermann et al 2005). In order to find the optimal quantity of zinc additive, different trials should be performed using controlled minimal media instead of rich LB (Outten and O'Halloran 2001).…”

Section: Protein Expressionmentioning

confidence: 99%

Recombinant Intrinsically Disordered Proteins for NMR: Tips and Tricks

Calçada

Korsak

Kozyreva

2015

Advances in Experimental Medicine and Biology

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The growing recognition of the several roles that intrinsically disordered proteins play in biology places an increasing importance on protein sample availability to allow the characterization of their structural and dynamic properties. The sample preparation is therefore the limiting step to allow any biophysical method being able to characterize the properties of an intrinsically disordered protein and to clarify the links between these properties and the associated biological functions. An increasing array of tools has been recruited to help prepare and characterize the structural and dynamic properties of disordered proteins. This chapter describes their sample preparation, covering the most common drawbacks/barriers usually found working in the laboratory bench. We want this chapter to be the bedside book of any scientist interested in preparing intrinsically disordered protein samples for further biophysical analysis.

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Effects of increased cellular zinc levels on gene and protein expression in HT-29 cells

Cited by 23 publications

References 49 publications

Zinc pyrithione induces cellular stress signaling and apoptosis in Hep-2 cervical tumor cells: the role of mitochondria and lysosomes

Zinc pyrithione induces cellular stress signaling and apoptosis in Hep-2 cervical tumor cells: the role of mitochondria and lysosomes

Zinc binding ligands and cellular zinc trafficking: Apo-metallothionein, glutathione, TPEN, proteomic zinc, and Zn-Sp1

Recombinant Intrinsically Disordered Proteins for NMR: Tips and Tricks

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