Expanding the Proteome of an RNA Virus by Phosphorylation of an Intrinsically Disordered Viral Protein

Cordek, Daniel G.; Croom-Perez, Tayler J.; Hwang, Jungwook; Hargittai, Michele R.S.; Subba-Reddy, Chennareddy V.; Han, Qingxia; Lodeiro, María F.; Ning, Gang; McCrory, Thomas S.; Arnold, Jamie J.; Koç, Hasan; Lindenbach, Brett D.; Showalter, Scott A.; Cameron, Craig Ε.

doi:10.1074/jbc.m114.589911

Cited by 18 publications

(19 citation statements)

References 60 publications

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“…Therefore, phosphorylation of IDPs appears to lead to a continuum of structure-promoting effects where PAGE4 is between amyloid precursor protein and 4E-BP2. Moreover, the structural ensemble shifts induced by posttranslational modifications seen here and elsewhere (57)(58)(59) are comparable with the conformational switching events in some marginally stable folded proteins in response to mutation or environmental triggers (60,61).…”

Section: Discussionmentioning

confidence: 52%

Phosphorylation-induced Conformational Ensemble Switching in an Intrinsically Disordered Cancer/Testis Antigen

Chen

Mooney

et al. 2015

Journal of Biological Chemistry

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Section: Discussionmentioning

confidence: 52%

Phosphorylation-induced Conformational Ensemble Switching in an Intrinsically Disordered Cancer/Testis Antigen

Chen

Mooney

et al. 2015

Journal of Biological Chemistry

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“…RNA viruses, such as HIV and Hepatitis C, utilize host kinases to phosphorylate viral proteins in order to increase the viral proteome to aid virus replication [54, 55]. Hepatitis C virus NS5A protein contains a phosphorylated threonine amongst proline-rich disordered residues that is crucial for replication center formation and production of virions [56]. In MNV-infected monocytes, NS1-2 is partially localized to the replication complex but it is not known what role it plays in replication [16].…”

Section: Resultsmentioning

confidence: 99%

Transcriptomic analysis of human norovirus NS1-2 protein highlights a multifunctional role in murine monocytes

et al. 2017

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BackgroundThe GII.4 Sydney 2012 strain of human norovirus (HuNoV) is a pandemic strain that is responsible for the majority of norovirus outbreaks in healthcare settings. The function of the non-structural (NS)1-2 protein from HuNoV is unknown.Results In silico analysis of human norovirus NS1-2 protein showed that it shares features with the murine NS1-2 protein, including a disordered region, a transmembrane domain and H-box and NC sequence motifs. The proteins also contain caspase cleavage and phosphorylation sites, indicating that processing and phosphorylation may be a conserved feature of norovirus NS1-2 proteins. In this study, RNA transcripts of human and murine norovirus full-length and the disordered region of NS1-2 were transfected into monocytes, and next generation sequencing was used to analyse the transcriptomic profile of cells expressing virus proteins. The profiles were then compared to the transcriptomic profile of MNV-infected cells.ConclusionsRNAseq analysis showed that NS1-2 proteins from human and murine noroviruses affect multiple immune systems (chemokine, cytokine, and Toll-like receptor signaling) and intracellular pathways (NFκB, MAPK, PI3K-Akt signaling) in murine monocytes. Comparison to the transcriptomic profile of MNV-infected cells indicated the pathways that NS1-2 may affect during norovirus infection.Electronic supplementary materialThe online version of this article (doi:10.1186/s12864-016-3417-4) contains supplementary material, which is available to authorized users.

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“…These high-resolution structures revealed a potential RNA binding groove (21). Together with biochemical data on RNA binding to NS5A (22), this led to the hypothesis that one function of NS5A is the translocation of the viral RNA from the site of replication to the site of particle assembly (23).…”

Section: Introductionmentioning

confidence: 96%

“…The main phosphorylation sites have been localized in the intrinsically disordered region of the protein (30). So far, protein kinases CK2, CK1a, PKA, and Plk1 have been shown-using kinase inhibitors and gene silencing-to be physiologically relevant for NS5A phosphorylation in vivo (21,27,31,32). To date, only a very few of the phosphorylation sites have been localized, and comprehensive data about eventual genotypic differences is lacking.…”

Section: Introductionmentioning

confidence: 97%

The Disordered Region of the HCV Protein NS5A: Conformational Dynamics, SH3 Binding, and Phosphorylation

Solyom

Schwarten

et al. 2015

Biophysical Journal

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Intrinsically disordered proteins (IDPs) perform their physiological role without possessing a well-defined three-dimensional structure. Still, residual structure and conformational dynamics of IDPs are crucial for the mechanisms underlying their functions. For example, regions of transient secondary structure are often involved in molecular recognition, with the structure being stabilized (or not) upon binding. Long-range interactions, on the other hand, determine the hydrodynamic radius of the IDP, and thus the distance over which the protein can catch binding partners via so-called fly-casting mechanisms. The modulation of long-range interactions also presents a convenient way of fine-tuning the protein's interaction network, by making binding sites more or less accessible. Here we studied, mainly by nuclear magnetic resonance spectroscopy, residual secondary structure and long-range interactions in nonstructural protein 5A (NS5A) from hepatitis C virus (HCV), a typical viral IDP with multiple functions during the viral life cycle. NS5A comprises an N-terminal folded domain, followed by a large (∼250-residue) disordered C-terminal part. Comparing nuclear magnetic resonance spectra of full-length NS5A with those of a protein construct composed of only the C-terminal residues 191-447 (NS5A-D2D3) allowed us to conclude that there is no significant interaction between the globular and disordered parts of NS5A. NS5A-D2D3, despite its overall high flexibility, shows a large extent of local residual (α-helical and β-turn) structure, as well as a network of electrostatic long-range interactions. Furthermore, we could demonstrate that these long-range interactions become modulated upon binding to the host protein Bin1, as well as after NS5A phosphorylation by CK2. As the charged peptide regions involved in these interactions are well conserved among the different HCV genotypes, these transient long-range interactions may be important for some of the functions of NS5A over the course of the HCV life cycle.

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Expanding the Proteome of an RNA Virus by Phosphorylation of an Intrinsically Disordered Viral Protein

Cited by 18 publications

References 60 publications

Phosphorylation-induced Conformational Ensemble Switching in an Intrinsically Disordered Cancer/Testis Antigen

Phosphorylation-induced Conformational Ensemble Switching in an Intrinsically Disordered Cancer/Testis Antigen

Transcriptomic analysis of human norovirus NS1-2 protein highlights a multifunctional role in murine monocytes

The Disordered Region of the HCV Protein NS5A: Conformational Dynamics, SH3 Binding, and Phosphorylation

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