scholarly journals The zinc-binding motif in tankyrases is required for the structural integrity and proper function of the catalytic domain

2021 ◽  
Author(s):  
Sven T Sowa ◽  
Lari Lehtiö
Keyword(s):  
Structural Integrity ◽  
Catalytic Domain ◽  
Zinc Binding ◽  
Structural Defects ◽  
Proper Function ◽  
Binding Motif ◽  
Acceptor Site ◽  
Structural Element ◽  
Zinc Binding Motif

Tankyrases are ADP-ribosylating enzymes that regulate many physiological processes in the cell and they are therefore possible drug targets for cancer and fibrotic diseases. The catalytic ADP-ribosyl-transferase domain of tankyrases contains a unique zinc-binding motif of unknown function. Recently, this motif was suggested to be involved in the catalytic activity of tankyrases. In this work, we set out to study the effect of the zinc-binding motif on activity, stability and structure of human tankyrases. We generated mutants of human TNKS1 and TNKS2 abolishing the zinc-binding capabilities and characterized the proteins biochemically and biophysically in vitro. We further generated a crystal structure of TNKS2, in which the zinc ion was oxidatively removed. Our work shows that the zinc-binding motif in tankyrases is a crucial structural element which is particularly important for the structural integrity of the acceptor site. While mutation of the motif rendered TNKS1 inactive likely due to introduction of major structural defects, the TNKS2 mutant remained active and displayed a different activity profile compared to the wild type.

Effect of mutations in a zinc-binding domain of yeast RNA polymerase C (III) on enzyme function and subunit association

1992 ◽  
Vol 12 (3) ◽  
pp. 1087-1095
Author(s):  
M Werner ◽  
S Hermann-Le Denmat ◽  
I Treich ◽  
A Sentenac ◽  
P Thuriaux
Keyword(s):  
Rna Polymerase ◽  
Zinc Binding ◽  
Enzyme Function ◽  
Directed Mutagenesis ◽  
Binding Motif ◽  
Zinc Ions ◽  
Amino Terminal ◽  
Zinc Binding Domain ◽  
Zinc Binding Motif

The conserved amino-terminal region of the largest subunit of yeast RNA polymerase C is capable of binding zinc ions in vitro. By oligonucleotide-directed mutagenesis, we show that the putative zinc-binding motif CX2CX6-12CXGHXGX24-37CX2C, present in the largest subunit of all eukaryotic and archaebacterial RNA polymerases, is essential for the function of RNA polymerase C. All mutations in the invariant cysteine and histidine residues conferred a lethal phenotype. We also obtained two conditional thermosensitive mutants affecting this region. One of these produced a form of RNA polymerase C which was thermosensitive and unstable in vitro. This instability was correlated with the loss of three of the subunits which are specific to RNA polymerase C: C82, C34, and C31.

scholarly journals Effect of mutations in a zinc-binding domain of yeast RNA polymerase C (III) on enzyme function and subunit association.

1992 ◽  
Vol 12 (3) ◽  
pp. 1087-1095 ◽  
Author(s):  
M Werner ◽  
S Hermann-Le Denmat ◽  
I Treich ◽  
A Sentenac ◽  
P Thuriaux
Keyword(s):  
Rna Polymerase ◽  
Zinc Binding ◽  
Enzyme Function ◽  
Directed Mutagenesis ◽  
Binding Motif ◽  
Zinc Ions ◽  
Amino Terminal ◽  
Zinc Binding Domain ◽  
Zinc Binding Motif

The conserved amino-terminal region of the largest subunit of yeast RNA polymerase C is capable of binding zinc ions in vitro. By oligonucleotide-directed mutagenesis, we show that the putative zinc-binding motif CX2CX6-12CXGHXGX24-37CX2C, present in the largest subunit of all eukaryotic and archaebacterial RNA polymerases, is essential for the function of RNA polymerase C. All mutations in the invariant cysteine and histidine residues conferred a lethal phenotype. We also obtained two conditional thermosensitive mutants affecting this region. One of these produced a form of RNA polymerase C which was thermosensitive and unstable in vitro. This instability was correlated with the loss of three of the subunits which are specific to RNA polymerase C: C82, C34, and C31.

scholarly journals Phosphonamidates are the first phosphorus-based zinc binding motif to show inhibition of β-class carbonic anhydrases from bacteria, fungi, and protozoa

2019 ◽  
Vol 35 (1) ◽  
pp. 59-64 ◽  
Author(s):  
Siham A. Alissa ◽  
Hanan A. Alghulikah ◽  
Zeid A. Alothman ◽  
Sameh M. Osman ◽  
Sonia Del Prete ◽  
...  
Keyword(s):  
Zinc Binding ◽  
Binding Motif ◽  
Carbonic Anhydrases ◽  
Zinc Binding Motif

Conformational dynamics and allosteric effect modulated by the unique zinc-binding motif in class IIa HDACs

2019 ◽  
Vol 21 (23) ◽  
pp. 12173-12183 ◽  
Author(s):  
Huawei Liu ◽  
Fan Zhang ◽  
Kai Wang ◽  
Xiaowen Tang ◽  
Ruibo Wu
Keyword(s):  
Histone Deacetylases ◽  
Conformational Dynamics ◽  
Zinc Binding ◽  
Binding Motif ◽  
Allosteric Effect ◽  
Class Iia Hdacs ◽  
Zinc Binding Motif

Class IIa histone deacetylases (HDACs) have been considered as potential targets for the treatment of several diseases.

scholarly journals Mutational analysis of the proteolytic domain of pregnancy-associated plasma protein-A (PAPP-A): classification as a metzincin

2001 ◽  
Vol 358 (2) ◽  
pp. 359-367 ◽  
Author(s):  
Henning B. BOLDT ◽  
Michael T. OVERGAARD ◽  
Lisbeth S. LAURSEN ◽  
Kathrin WEYER ◽  
Lars SOTTRUP-JENSEN ◽  
...  
Keyword(s):  
Plasma Protein ◽  
Active Site ◽  
Mammalian Cells ◽  
Mutational Analysis ◽  
Protein A ◽  
Residual Activity ◽  
Zinc Binding ◽  
Binding Motif ◽  
Dependent Manner ◽  
Zinc Binding Motif

The bioavailability of insulin-like growth factor (IGF)-I and -II is controlled by six IGF-binding proteins (IGFBPs 1–6). Bound IGF is not active, but proteolytic cleavage of the binding protein causes release of IGF. Pregnancy-associated plasma protein-A (PAPP-A) has recently been found to cleave IGFBP-4 in an IGF-dependent manner. To experimentally support the hypothesis that PAPP-A belongs to the metzincin superfamily of metalloproteinases, all containing the elongated zinc-binding motif HEXXHXXGXXH (His-482–His-492 in PAPP-A), we expressed mutants of PAPP-A in mammalian cells. Substitution of Glu-483 with Ala causes a complete loss of activity, defining this motif as part of the active site of PAPP-A. Interestingly, a mutant with Glu-483 replaced by Gln shows residual activity. Known metzincin structures contain a so-called Met-turn, whose strictly conserved Met residue is thought to interact directly with residues of the active site. By further mutagenesis we provide experimental evidence that Met-556 of PAPP-A, 63 residues from the zinc-binding motif, is located in a Met-turn of PAPP-A. Our hypothesis is also supported by secondary-structure prediction, and the ability of a 55-residue deletion mutant (d[S498-Y552]) to express and retain antigenecity. However, because PAPP-A differs in the features defining the individual established metzincin families, we suggest that PAPP-A belongs to a separate family. We also found that PAPP-A can undergo autocleavage, and that autocleaved PAPP-A is inactive. A lack of unifying elements in the sequences around the found cleavage sites of PAPP-A and a variant suggests steric regulation of substrate specificity.

COP1, an arabidopsis regulatory gene, encodes a protein with both a zinc-binding motif and a Gβ homologous domain

Cell ◽  
1992 ◽  
Vol 71 (5) ◽  
pp. 791-801 ◽  
Author(s):  
Xing-Wang Deng ◽  
Minami Matsui ◽  
Ning Wei ◽  
Doris Wagner ◽  
Angela M. Chu ◽  
...  
Keyword(s):  
Regulatory Gene ◽  
Zinc Binding ◽  
Binding Motif ◽  
Zinc Binding Motif

scholarly journals The zinc-binding motif of human RECQ5β suppresses the intrinsic strand-annealing activity of its DExH helicase domain and is essential for the helicase activity of the enzyme

2008 ◽  
Vol 412 (3) ◽  
pp. 425-433 ◽  
Author(s):  
Hua Ren ◽  
Shuo-Xing Dou ◽  
Xing-Dong Zhang ◽  
Peng-Ye Wang ◽  
Radhakrishnan Kanagaraj ◽  
...  
Keyword(s):  
Structural Similarity ◽  
Zinc Binding ◽  
Binding Motif ◽  
Dna Unwinding ◽  
Helicase Domain ◽  
The Novel ◽  
Quantitative Measurements ◽  
Strand Annealing ◽  
Zinc Binding Motif

RecQ family helicases, functioning as caretakers of genomic integrity, contain a zinc-binding motif which is highly conserved among these helicases, but does not have a substantial structural similarity with any other known zinc-finger folds. In the present study, we show that a truncated variant of the human RECQ5β helicase comprised of the conserved helicase domain only, a splice variant named RECQ5α, possesses neither ATPase nor DNA-unwinding activities, but surprisingly displays a strong strand-annealing activity. In contrast, fragments of RECQ5β including the intact zinc-binding motif, which is located immediately downstream of the helicase domain, exhibit much reduced strand-annealing activity but are proficient in DNA unwinding. Quantitative measurements indicate that the regulatory role of the zinc-binding motif is achieved by enhancing the DNA-binding affinity of the enzyme. The novel intramolecular modulation of RECQ5β catalytic activity mediated by the zinc-binding motif may represent a universal regulation mode for all RecQ family helicases.

scholarly journals Redox Switch of Hsp33 Has a Novel Zinc-binding Motif

2000 ◽  
Vol 275 (49) ◽  
pp. 38302-38310 ◽  
Author(s):  
Ursula Jakob ◽  
Markus Eser ◽  
James C. A. Bardwell
Keyword(s):  
Zinc Binding ◽  
Binding Motif ◽  
Redox Switch ◽  
Zinc Binding Motif
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