The Leishmania donovani IMPACT-like protein possesses non-specific nuclease activity

Rao, Shilpa; Shukla, Ekta; Bhatia, Varsha; Lohiya, Bharat; Gaikwad, Sushama M.; Kar, Anita; Pal, Jayanta K.

doi:10.1016/j.ijbiomac.2018.07.175

Cited by 2 publications

(1 citation statement)

References 34 publications

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“…We can speculate, that if these motifs are involved in binding yet‐to‐be‐discovered binding partners, their effect on interdomain interactions within Yih1, is likely to be via entropically based mechanisms, as is increasingly found in allosteric regulation [51–53]. The most likely binding partner for this region is DNA or RNA, consistent with the recently discovered nuclease activity of IMPACT proteins [54,55]. The nuclease activity of the fungus Chaetomium thermophilum IMPACT homolog [54] and the predicted nucleic acid binding interface of YigZ protein [36] can be mapped to this region (Fig.…”

Section: Discussionsupporting

confidence: 63%

Experimentally based structural model of Yih1 provides insight into its function in controlling the key translational regulator Gcn2

Harjes

Jameson

et al. 2020

FEBS Letters

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Yeast impact homolog 1 (Yih1), or IMPACT in mammals, is part of a conserved regulatory module controlling the activity of General Control Nonderepressible 2 (Gcn2), a protein kinase that regulates protein synthesis. Yih1/IMPACT is implicated not only in many essential cellular processes, such as neuronal development, immune system regulation and the cell cycle, but also in cancer. Gcn2 must bind to Gcn1 in order to impair the initiation of protein translation. Yih1 hinders this key Gcn1‐Gcn2 interaction by binding to Gcn1, thus preventing Gcn2‐mediated inhibition of protein synthesis. Here, we solved the structures of the two domains of Saccharomyces cerevisiae Yih1 separately using Nuclear Magnetic Resonance and determined the relative positions of the two domains using a range of biophysical methods. Our findings support a compact structural model of Yih1 in which the residues required for Gcn1 binding are buried in the interface. This model strongly implies that Yih1 undergoes a large conformational rearrangement from a latent closed state to a primed open state to bind Gcn1. Our study provides structural insight into the interactions of Yih1 with partner molecules.

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Section: Discussionsupporting

confidence: 63%

Experimentally based structural model of Yih1 provides insight into its function in controlling the key translational regulator Gcn2

Harjes

Jameson

et al. 2020

FEBS Letters

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Decoding Essential Amino Acid Residues in the Substrate Groove of a Non-Specific Nuclease from Pseudomonas syringae

et al. 2019

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Non-specific nucleases (NSN) are of interest for biotechnological applications, including industrial downstream processing of crude protein extracts or cell-sorting approaches in microfabricated channels. Bacterial nucleases belonging to the superfamily of phospholipase D (PLD) are featured for their ability to catalyze the hydrolysis of nucleic acids in a metal-ion-independent manner. In order to gain a deeper insight into the composition of the substrate groove of a NSN from Pseudomonas syringae, semi-rational mutagenesis based on a structure homology model was applied to identify amino acid residues on the protein’s surface adjacent to the catalytic region. A collection of 12 mutant enzymes each with a substitution to a positively charged amino acid (arginine or lysine) was produced in recombinant form and biochemically characterized. Mutations in close proximity to the catalytic region (inner ring) either dramatically impaired or completely abolished the enzymatic performance, while amino acid residues located at the border of the substrate groove (outer ring) only had limited or no effects. A K119R substitution mutant displayed a relative turnover rate of 112% compared to the original nuclease. In conclusion, the well-defined outer ring of the substrate groove is a potential target for modulation of the enzymatic performance of NSNs belonging to the PLD superfamily.

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The Leishmania donovani IMPACT-like protein possesses non-specific nuclease activity

Cited by 2 publications

References 34 publications

Experimentally based structural model of Yih1 provides insight into its function in controlling the key translational regulator Gcn2

Experimentally based structural model of Yih1 provides insight into its function in controlling the key translational regulator Gcn2

Decoding Essential Amino Acid Residues in the Substrate Groove of a Non-Specific Nuclease from Pseudomonas syringae

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