Cu(I) Disrupts the Structure and Function of the Nonclassical Zinc Finger Protein Tristetraprolin (TTP)

Shimberg, Geoffrey D.; Ok, Kiwon; Neu, Heather M.; Splan, Kathryn E.; Michel, Sarah L. J.

doi:10.1021/acs.inorgchem.7b00125

Cited by 27 publications

(54 citation statements)

References 93 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…[18] Excess Au III terpy (5 equiv) was added to as olutiono f Zn II 2 -TTP-2D. [18] Excess Au III terpy (5 equiv) was added to as olutiono f Zn II 2 -TTP-2D.…”

Section: Directt Argeting Of Zn II 2 -Ttp-2d With Goldmentioning

confidence: 63%

“…[1a, 3] As such, it is common to over-express just the ZF domains for study. [7,18] We chose Au III terpy as the gold complex for study,a nd synthesized the complex using published protocols (Figure 1C). [7,18] We chose Au III terpy as the gold complex for study,a nd synthesized the complex using published protocols (Figure 1C).…”

Section: Resultsmentioning

confidence: 99%

“…[7,18] Briefly,t he plasmid was transformed into E. Coli BL21 (DE3)-competent cells (Novagen) and grown in 1L of Luria-Bertani (LB) medium containing 100 mgmL À1 ampicillin and 100 mm ZnCl 2 at 37 8Cu ntil mid-log phase (% OD 600 of 0.6-0.8). [7,18] Briefly,t he plasmid was transformed into E. Coli BL21 (DE3)-competent cells (Novagen) and grown in 1L of Luria-Bertani (LB) medium containing 100 mgmL À1 ampicillin and 100 mm ZnCl 2 at 37 8Cu ntil mid-log phase (% OD 600 of 0.6-0.8).…”

Section: Ttp-2d Overexpression and Purificationmentioning

confidence: 99%

“…[1a, 3] We took this approacha nd isolated ac onstruct of TTP called TTP-2D, which contains the protein'st wo CCCH domains.W eh ave previously reported the cloning, overexpression and purification of TTP-2D, and demonstrated that it is highly soluble, binds and folds aroundz inc, and functions (selectively recognize AU-rich mRNA targets)( Figure 1A,B). [7,18] We chose Au III terpy as the gold complex for study,a nd synthesized the complex using published protocols (Figure 1C). [13b, 19] Au III terpy is soluble in aqueous solution and it exhibits rich spectroscopy (charge-transfer bands) that are sensitive to metal coordination, allowing for reactivity to be followed optically.M oreover,t here are severalr ecent reports of the interaction of gold complexes,i ncluding Au III terpy,w ith ZF peptides, including Cys 4 "zinc ribbon"s ites, [13b] Cys 2 HisCys "zinc knuckle", [12,14,20] Cys 2 HisCys( PARP-1), [13,21] andC ys 2 His 2 classical ZF, [12a,c, 14, 22] and consensus peptide1(Cys 2 His 2 ).…”

Section: Choice Of Thet Tp and Gold Complexmentioning

confidence: 99%

See 3 more Smart Citations

Role of Gold in Inflammation and Tristetraprolin Activity

Neu

et al. 2020

Chemistry A European J

Self Cite

View full text Add to dashboard Cite

The zinc finger protein tristetraprolin (TTP) regulates inflammation by downregulatingc ytokinem RNAs. Misregulation resultsi na rthritis, sepsis and cancer,a nd there is an interesti nm odulating TTP activity with exogenous agents. Gold hasa nti-inflammatory properties and has recently been shown to modulate the signaling pathway that produces TTP,s uggesting that TTP may be at arget of gold. The reactivity of [Au III (terpy)Cl]Cl 2 with TTPw as investigated by UV/Vis spectroscopy,s pin-filter inductively coupled plasma mass spectrometry,X -ray absorption spectroscopy and native electrospray ionization mass spectrometry.A u III was found to replace zinc in the protein active site in the reduced Au I form, with the Au I ion coordinated to two cysteine residues in al inear geometry.T he replacement of Zn II with Au I resultsi nl oss of both secondary structure and RNA binding function. In contrast, when Zn II TTP is boundt oi ts RNA target, no replacement of Zn II with Au I is observed, even in the presence of excessA u III terpy.T his discovery of differential reactivity of gold with TTP versus TTP/RNA offers ap otentials trategy for selectivet argeting with gold complexest oc ontroli nflammation.Supporting information and the ORCID identification number(s) for the author(s) of this articlecan be found under: https://doi.org/10.1002/chem.201904837.Scheme1.Cartoon diagram of role of TTP in TNF-a signaling. TTP (in yellow) regulates TNF-a by binding to and degrading its mRNA as part of the NF-kBs ignaling pathway.

show abstract

“…[18] Excess Au III terpy (5 equiv) was added to as olutiono f Zn II 2 -TTP-2D. [18] Excess Au III terpy (5 equiv) was added to as olutiono f Zn II 2 -TTP-2D.…”

Section: Directt Argeting Of Zn II 2 -Ttp-2d With Goldmentioning

confidence: 63%

Section: Resultsmentioning

confidence: 99%

Section: Ttp-2d Overexpression and Purificationmentioning

confidence: 99%

Section: Choice Of Thet Tp and Gold Complexmentioning

confidence: 99%

See 2 more Smart Citations

Role of Gold in Inflammation and Tristetraprolin Activity

Neu

et al. 2020

Chemistry A European J

Self Cite

View full text Add to dashboard Cite

show abstract

“…Whereas divalent ions, such as Zn 2+ , positively influence phase separation [ 12 ], others (e.g., Mg 2+ , Ca 2+ ) negatively influence PrLD–RNA interactions and reduce LLPS, as does the metal chelator EDTA [ 128 ]. Although Zn 2+ is the presumed metal binding ZnFs, ZnFs also coordinate numerous other metal ions (reviewed in [ 129 ]). As the third most abundant metal following iron (Fe) and Zn in eukaryotic cells, copper (Cu) is of particular interest because its binding to classical and non-classical ZnFs, including HIV-1 NCp7, is thermodynamically favored over the binding of other metals, including Zn 2+ [ 130 ].…”

Section: Resultsmentioning

confidence: 99%

Zinc and Copper Ions Differentially Regulate Prion-Like Phase Separation Dynamics of Pan-Virus Nucleocapsid Biomolecular Condensates

Monette

Mouland

2020

Viruses

View full text Add to dashboard Cite

Liquid-liquid phase separation (LLPS) is a rapidly growing research focus due to numerous demonstrations that many cellular proteins phase-separate to form biomolecular condensates (BMCs) that nucleate membraneless organelles (MLOs). A growing repertoire of mechanisms supporting BMC formation, composition, dynamics, and functions are becoming elucidated. BMCs are now appreciated as required for several steps of gene regulation, while their deregulation promotes pathological aggregates, such as stress granules (SGs) and insoluble irreversible plaques that are hallmarks of neurodegenerative diseases. Treatment of BMC-related diseases will greatly benefit from identification of therapeutics preventing pathological aggregates while sparing BMCs required for cellular functions. Numerous viruses that block SG assembly also utilize or engineer BMCs for their replication. While BMC formation first depends on prion-like disordered protein domains (PrLDs), metal ion-controlled RNA-binding domains (RBDs) also orchestrate their formation. Virus replication and viral genomic RNA (vRNA) packaging dynamics involving nucleocapsid (NC) proteins and their orthologs rely on Zinc (Zn) availability, while virus morphology and infectivity are negatively influenced by excess Copper (Cu). While virus infections modify physiological metal homeostasis towards an increased copper to zinc ratio (Cu/Zn), how and why they do this remains elusive. Following our recent finding that pan-retroviruses employ Zn for NC-mediated LLPS for virus assembly, we present a pan-virus bioinformatics and literature meta-analysis study identifying metal-based mechanisms linking virus-induced BMCs to neurodegenerative disease processes. We discover that conserved degree and placement of PrLDs juxtaposing metal-regulated RBDs are associated with disease-causing prion-like proteins and are common features of viral proteins responsible for virus capsid assembly and structure. Virus infections both modulate gene expression of metalloproteins and interfere with metal homeostasis, representing an additional virus strategy impeding physiological and cellular antiviral responses. Our analyses reveal that metal-coordinated virus NC protein PrLDs initiate LLPS that nucleate pan-virus assembly and contribute to their persistence as cell-free infectious aerosol droplets. Virus aerosol droplets and insoluble neurological disease aggregates should be eliminated by physiological or environmental metals that outcompete PrLD-bound metals. While environmental metals can control virus spreading via aerosol droplets, therapeutic interference with metals or metalloproteins represent additional attractive avenues against pan-virus infection and virus-exacerbated neurological diseases.

show abstract

Targeting Zinc Finger Proteins with Exogenous Metals and Molecules: Lessons Learned from Tristetraprolin, a CCCH type Zinc Finger

Filipović

Michel

2021

Eur J Inorg Chem

Self Cite

View full text Add to dashboard Cite

ZF proteins are ubiquitous eukaryotic proteins that play important roles in gene regulation. ZFs contain small domains made up of a combination of four cysteine and histidine residues and are classified on the basis of the identity of these residues and their spacing. One emerging class of ZFs are the Cys3His (or CCCH) class of ZFs. These ZFs play key roles in regulating RNA. In this minireview, an overview of the CCCH class of ZFs, with a focus on tristetraprolin (TTP), is provided. TTP regulates inflammation by controlling cytokine mRNAs, and there is an interest in modulating TTP activity to control inflammation. Two methods to control TTP activity are to target with exogenous metals (a “metals in medicine” approach) or to target with endogenous signaling molecules. Work that has been done to target TTP with Fe, Cu, Cd, and Au as well as with H2S is reviewed. This includes attention to new methods that have been developed to monitor metal exchange with the spectroscopically silent ZnII including native electro‐spray ionization mass spectrometry (ESI‐MS), spin‐filter inductively coupled plasma mass spectrometry (ICP‐MS), and cryo‐electro‐spray mass spectrometry (CSI‐MS); along with fluorescence anisotropy (FA) to follow RNA binding.

show abstract

Cu(I) Disrupts the Structure and Function of the Nonclassical Zinc Finger Protein Tristetraprolin (TTP)

Cited by 27 publications

References 93 publications

Role of Gold in Inflammation and Tristetraprolin Activity

Role of Gold in Inflammation and Tristetraprolin Activity

Zinc and Copper Ions Differentially Regulate Prion-Like Phase Separation Dynamics of Pan-Virus Nucleocapsid Biomolecular Condensates

Targeting Zinc Finger Proteins with Exogenous Metals and Molecules: Lessons Learned from Tristetraprolin, a CCCH type Zinc Finger

Contact Info

Product

Resources

About