Probing the Interaction between U24 and the SH3 Domain of Fyn Tyrosine Kinase

Sang, Yurou; Tait, Andrew R.; Scott, Walter R. P.; Creagh, A. Louise; Kumar, Prashant; Haynes, Charles A.; Straus, Suzana K.

doi:10.1021/bi500945x

Cited by 7 publications

(8 citation statements)

References 46 publications

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“…In the future, it will be particularly challenging, but necessary, to probe spectroscopically how membrane-associated 18.5 kDa MBP interacts with SH3–domain containing proteins and how combinatorial phosphorylation events can modulate these multifaceted in situ interactions. It is important to understand these events further because interference in these normal associations during brain development has been suggested to result potentially in myelin instability and subsequent pathologies [ 10 , 82 – 86 ].…”

Section: Discussionmentioning

confidence: 99%

The proline-rich region of 18.5 kDa myelin basic protein binds to the SH3-domain of Fyn tyrosine kinase with the aid of an upstream segment to form a dynamic complexin vitro

et al. 2014

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The intrinsically disordered 18.5 kDa classic isoform of MBP (myelin basic protein) interacts with Fyn kinase during oligodendrocyte development and myelination. It does so primarily via a central proline-rich SH3 (Src homology 3) ligand (T92–R104, murine 18.5 kDa MBP sequence numbering) that is part of a molecular switch due to its high degree of conservation and modification by MAP (mitogen-activated protein) and other kinases, especially at residues T92 and T95. Here, we show using co-transfection experiments of an early developmental oligodendroglial cell line (N19) that an MBP segment upstream of the primary ligand is involved in MBP–Fyn–SH3 association in cellula. Using solution NMR spectroscopy in vitro, we define this segment to comprise MBP residues (T62–L68), and demonstrate further that residues (V83–P93) are the predominant SH3-target, assessed by the degree of chemical shift change upon titration. We show by chemical shift index analysis that there is no formation of local poly-proline type II structure in the proline-rich segment upon binding, and by NOE (nuclear Overhauser effect) and relaxation measurements that MBP remains dynamic even while complexed with Fyn–SH3. The association is a new example first of a non-canonical SH3-domain interaction and second of a fuzzy MBP complex.

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Section: Discussionmentioning

confidence: 99%

The proline-rich region of 18.5 kDa myelin basic protein binds to the SH3-domain of Fyn tyrosine kinase with the aid of an upstream segment to form a dynamic complexin vitro

et al. 2014

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“…The WW domains mentioned above were fused to the C-terminus of GST protein individually. Recombinant full length U24-6A and U24-7 protein were prepared as previously described1438.…”

Section: Resultsmentioning

confidence: 99%

“…U24-6A and U24-7 were prepared as described previously14. The resin was incubated with 600 μL U24-6A and U24-7 protein stock solution in pull down buffer at 4 °C.…”

Section: Methodsmentioning

confidence: 99%

“…( a ) The numbering of MBP is based on the human 18.5 kDa classic MBP (Note that in ref. 14, the murine numbering 90–104 was used). The identical segment between U24-6A and MBP, PRTPPPS, is marked with yellow shading.…”

Section: Figurementioning

confidence: 99%

“…Recent work has demonstrated that U24-6A can be phosphorylated by MAPK, albeit less effectively than MBP13. It has also been shown that U24-6A can associate with Fyn SH3, but again does so more weakly than MBP ( K D = 5 mM for U24-6A14 versus 4-8 μM for MBP15). These findings suggested that although U24-6A may mimic MBP, it most likely does not interfere directly with MBP function.…”

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confidence: 99%

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U24 from Roseolovirus interacts strongly with Nedd4 WW Domains

Sang

Zhang

Scott

et al. 2017

Sci Rep

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U24 is a protein found in both roseoloviruses Human Herpes Virus type 6 and 7 (HHV-6 and HHV-7), with an N-terminus that is rich in prolines (PY motif in both HHV-6A and 7; PxxP motif in HHV-6A). Previous work has shown that the interaction between U24 and WW domains is important for endocytic recycling of T-cell receptors, but a cognate ligand was never identified. In this contribution, data was obtained from pull-downs, ITC, NMR and molecular dynamics simulations to show that a specific interaction exists between U24 and Nedd4 WW domains. ITC experiments were also carried out for U24 from HHV-6A phosphorylated at Thr6 (pU24-6A) and a peptide containing the PY motif from Nogo-A, a protein implicated in both the initial inflammatory and the neurodegenerative phases of multiple sclerosis (MS). The results suggest that phosphorylation of U24 from HHV-6A may be crucial for its potential role in MS.Roseoloviruses, which include human herpes virus type 6A (HHV-6A), HHV-6B and HHV-7, are ubiquitous and highly prevalent. They have been found to be implicated in several neurological diseases, including encephalitis, epilepsy, and multiple sclerosis (MS) [1][2][3][4] . MS is a neurological inflammatory disease in the central nervous system (CNS). The exact cause of MS is unknown, but it has been suggested that one or more viruses may be possible triggers 2,5,6 . U24 is a putative tail-anchored membrane protein that is unique to roseoloviruses 7 . It is expressed in infected cells during the early stages of viral infection 8 . Although the exact function of U24 has not been identified, a number of studies have demonstrated a potential link between U24 and MS 9,10 .One possible way in which U24 may be implicated in MS is through the mimicry of myelin basic protein (MBP) because the two proteins share a seven amino acid stretch that contains a PxxP motif (Fig. 1a). This segment is essential for MBP to participate in Fyn-mediated signalling pathways by a direct but non-canonical association with the Fyn-SH3 domain 11 . Disruption of these signalling pathways has a large impact on oligodendrocyte differentiation and maturation, processes that are perturbed in MS patients. Within this PxxP motif and just before it are two threonine residues (T95 and T98), which can be phosphorylated. Phosphorylation has been shown to affect the local structure of MBP and its disposition on the membrane surface 12 . U24-6A shares with MBP both the PxxP motif and a threonine at an analogous position to T98 (Fig. 1a). Recent work has demonstrated that U24-6A can be phosphorylated by MAPK, albeit less effectively than MBP 13 . It has also been shown that U24-6A can associate with Fyn SH3, but again does so more weakly than MBP (K D = 5 mM for U24-6A 14 versus 4-8 μ M for MBP 15 ). These findings suggested that although U24-6A may mimic MBP, it most likely does not interfere directly with MBP function.Within the proline rich region of U24 is a PY motif that is conserved in both HHV-6A and HHV-7 (Fig. 1a). This motif was found to be responsible for ...

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Multiscale Analysis of Independent Alzheimer’s Cohorts Finds Disruption of Molecular, Genetic, and Clinical Networks by Human Herpesvirus

Readhead

Haure‐Mirande

Funk

et al. 2018

Neuron

572

475

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Investigators have long suspected that pathogenic microbes might contribute to the onset and progression of Alzheimer's disease (AD) although definitive evidence has not been presented. Whether such findings represent a causal contribution, or reflect opportunistic passengers of neurodegeneration, is also difficult to resolve. We constructed multiscale networks of the late-onset AD-associated virome, integrating genomic, transcriptomic, proteomic, and histopathological data across four brain regions from human post-mortem tissue. We observed increased human herpesvirus 6A (HHV-6A) and human herpesvirus 7 (HHV-7) from subjects with AD compared with controls. These results were replicated in two additional, independent and geographically dispersed cohorts. We observed regulatory relationships linking viral abundance and modulators of APP metabolism, including induction of APBB2, APPBP2, BIN1, BACE1, CLU, PICALM, and PSEN1 by HHV-6A. This study elucidates networks linking molecular, clinical, and neuropathological features with viral activity and is consistent with viral activity constituting a general feature of AD.

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Probing the Interaction between U24 and the SH3 Domain of Fyn Tyrosine Kinase

Cited by 7 publications

References 46 publications

The proline-rich region of 18.5 kDa myelin basic protein binds to the SH3-domain of Fyn tyrosine kinase with the aid of an upstream segment to form a dynamic complexin vitro

The proline-rich region of 18.5 kDa myelin basic protein binds to the SH3-domain of Fyn tyrosine kinase with the aid of an upstream segment to form a dynamic complexin vitro

U24 from Roseolovirus interacts strongly with Nedd4 WW Domains

Multiscale Analysis of Independent Alzheimer’s Cohorts Finds Disruption of Molecular, Genetic, and Clinical Networks by Human Herpesvirus

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