Structural and biophysical analysis of sequence insertions in the Venezuelan Equine Encephalitis Virus macro domain

Guillén, Jaime; Lichière, Julie; Rabah, Nadia; Beitzel, Brett; Canard, Bruno; Coutard, Bruno

doi:10.1016/j.virusres.2015.02.018

Cited by 2 publications

(3 citation statements)

References 42 publications

(47 reference statements)

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“…The displacement of the b3-a1 loop could also explain the lower DT m upon OAADPr binding at different concentrations (figure 5c inset), and the slight increase in K m (table 2) compared with WT. Collectively, these results suggest a dual role for residue D40, a catalytic role as the main activator of the catalytic water and a novel structural function in the organization of the b3-a1 loop, which is essential for maintaining the proper architecture of the binding site for ADPr and related molecules, as has been previously described [9,26,30].…”

Section: Mutant D40a Reveals An Additional Structural Role For Residue D40supporting

confidence: 70%

Structural and functional analysis ofOceanobacillus iheyensismacrodomain reveals a network of waters involved in substrate binding and catalysis

et al. 2017

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Macrodomains are ubiquitous conserved domains that bind or transform ADP-ribose (ADPr) metabolites. In humans, they are involved in transcription, X-chromosome inactivation, neurodegeneration and modulating PARP1 signalling, making them potential targets for therapeutic agents. Unfortunately, some aspects related to the substrate binding and catalysis of MacroD-like macrodomains still remain unclear, since mutation of the proposed catalytic aspartate does not completely abolish enzyme activity. Here, we present a functional and structural characterization of a macrodomain from the extremely halotolerant and alkaliphilic bacterium Oceanobacillus iheyensis (OiMacroD), related to hMacroD1/hMacroD2, shedding light on substrate binding and catalysis. The crystal structures of D40A, N30A and G37V mutants, and those with MES, ADPr and ADP bound, allowed us to identify five fixed water molecules that play a significant role in substrate binding. Closure of the β6–α4 loop is revealed as essential not only for pyrophosphate recognition, but also for distal ribose orientation. In addition, a novel structural role for residue D40 is identified. Furthermore, it is revealed that OiMacroD not only catalyses the hydrolysis of O-acetyl-ADP-ribose but also reverses protein mono-ADP-ribosylation. Finally, mutant G37V supports the participation of a substrate-coordinated water molecule in catalysis that helps to select the proper substrate conformation.

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Section: Mutant D40a Reveals An Additional Structural Role For Residue D40supporting

confidence: 70%

Structural and functional analysis ofOceanobacillus iheyensismacrodomain reveals a network of waters involved in substrate binding and catalysis

et al. 2017

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“…Consequently, F114 also moves to interact with the same ribose. Mutational insertions in the Q28-A36 loop can conditionally alter virus replication and, at the protein scale, destabilize the macro domain and partly affect its ADPr binding property (Beitzel et al, 2010;Guillén et al, 2015), highlighting the role this loop in the transition mechanism required for the accommodation of ADPr.…”

Section: Dynamic and Exchange Properties Of The Veev Macro Domain (Apmentioning

confidence: 99%

“…Such outbreaks in Venezuela and Colombia in 1995, involving an estimated 100,000 human cases, indicate that VEEV poses a serious public health problem. It is worth mentioning that since 1995 there has not been any outbreak of the virus (Guillén et al, 2015;Morrison et al, 2008), however epidemics indicate that VEEV still represents a serious public health concern (Aguilar et al, 2011). So far, no efficient vaccines or drugs are licensed for the treatment of alphavirus infection (Chattopadhyay et al, 2013).…”

Section: Introductionmentioning

confidence: 99%

Conformational plasticity of the VEEV macro domain is important for binding of ADP-ribose

Makrynitsa

Ntonti

Marousis

et al. 2019

Journal of Structural Biology

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A B S T R A C TVenezuelan equine encephalitis virus (VEEV) is a new world alphavirus which can be involved in several central nervous system disorders such as encephalitis and meningitis. The VEEV genome codes for 4 non-structural proteins (nsP), of which nsP3 contains a Macro domain. Macro domains (MD) can be found as stand-alone proteins or embedded within larger proteins in viruses, bacteria and eukaryotes. Their most common feature is the binding of ADP-ribose (ADPr), while several macro domains act as ribosylation writers, erasers or readers. Alphavirus MD erase ribosylation but their precise contribution in viral replication is still under investigation. NMR-driven titration experiments of ADPr in solution with the VEEV macro domain (in apo-and complex state) show that it adopts a suitable conformation for ADPr binding. Specific experiments indicate that the flexibility of the loops β5-α3 and α3-β6 is critical for formation of the complex and assists a wrapping mechanism for ADPr binding. Furthermore, along with this sequence of events, the VEEV MD undergoes a conformational exchange process between the apo state and a low-populated "dark" conformational state.

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Structural and biophysical analysis of sequence insertions in the Venezuelan Equine Encephalitis Virus macro domain

Cited by 2 publications

References 42 publications

Structural and functional analysis ofOceanobacillus iheyensismacrodomain reveals a network of waters involved in substrate binding and catalysis

Structural and functional analysis ofOceanobacillus iheyensismacrodomain reveals a network of waters involved in substrate binding and catalysis

Conformational plasticity of the VEEV macro domain is important for binding of ADP-ribose

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