NMR Studies of the Phosphorylation Motif of the HIV-1 Protein Vpu Bound to the F-Box Protein β-TrCP†

Biochemistry ◽  
2003 ◽  
Vol 42 (50) ◽  
pp. 14741-14751 ◽  
Author(s):  
Gaël Coadou ◽  
Josyane Gharbi-Benarous ◽  
Simon Megy ◽  
Gildas Bertho ◽  
Nathalie Evrard-Todeschi ◽  
...  
Keyword(s):  
Nmr Studies ◽  
Phosphorylation Motif ◽  
F Box Protein ◽  
Hiv 1

NMR studies for identifying phosphopeptide ligands of the HIV-1 protein Vpu binding to the F-box protein β-TrCP

Peptides ◽  
2006 ◽  
Vol 27 (1) ◽  
pp. 194-210 ◽  
Author(s):  
Nathalie Evrard-Todeschi ◽  
Josyane Gharbi-Benarous ◽  
Gildas Bertho ◽  
Gaël Coadou ◽  
Simon Megy ◽  
...  
Keyword(s):  
Nmr Studies ◽  
F Box Protein ◽  
Hiv 1

scholarly journals Solid-State NMR Studies of HIV-1 Capsid Protein Assemblies

2010 ◽  
Vol 132 (6) ◽  
pp. 1976-1987 ◽  
Author(s):  
Yun Han ◽  
Jinwoo Ahn ◽  
Jason Concel ◽  
In-Ja L. Byeon ◽  
Angela M. Gronenborn ◽  
...  
Keyword(s):  
Solid State ◽  
Capsid Protein ◽  
Solid State Nmr ◽  
Protein Assemblies ◽  
Nmr Studies ◽  
Hiv 1

scholarly journals Ubiquitylation of Cdk9 by Skp2 Facilitates Optimal Tat Transactivation

2005 ◽  
Vol 79 (17) ◽  
pp. 11135-11141 ◽  
Author(s):  
Matjaz Barboric ◽  
Fan Zhang ◽  
Mojca Besenicar ◽  
Ana Plemenitas ◽  
B. Matija Peterlin
Keyword(s):  
Rna Polymerase Ii ◽  
Elongation Factor ◽  
Transcriptional Elongation ◽  
Cyclin Dependent Kinase ◽  
Primary Mouse ◽  
Immunodeficiency Virus ◽  
F Box Protein ◽  
Hiv 1

ABSTRACT By recruiting the positive transcriptional elongation factor b (P-TEFb) to paused RNA polymerase II, the transactivator Tat stimulates transcriptional elongation of the human immunodeficiency virus type 1 (HIV-1) genome. We found that cyclin-dependent kinase 9 (Cdk9), the catalytic subunit of P-TEFb, is ubiquitylated in vivo. This ubiquitylation depended on the Skp1/Cul1/F-box protein E3 ubiquitin ligase Skp2. Likewise, Tat required Skp2 since its transactivation of the HIV-1 long terminal repeat decreased in primary mouse embryonic fibroblasts, which lacked Skp2. The ubiquitylation of Cdk9 by Skp2 facilitated the formation of the ternary complex between P-TEFb, Tat, and transactivation response element. Thus, our findings underscore the requirement of ubiquitylation for the coactivator function in regulating HIV-1 transcriptional elongation.

scholarly journals 19F NMR Studies of Cyclophilin a and its Interaction with HIV-1 Capsid

2020 ◽  
Vol 118 (3) ◽  
pp. 503a
Author(s):  
Manman Lu ◽  
Tatyana E. Polenova ◽  
Angela M. Gronenborn
Keyword(s):  
Cyclophilin A ◽  
19F Nmr ◽  
Nmr Studies ◽  
Hiv 1

scholarly journals Identification of the initial nucleocapsid recognition element in the HIV-1 RNA packaging signal

2020 ◽  
Vol 117 (30) ◽  
pp. 17737-17746 ◽  
Author(s):  
Pengfei Ding ◽  
Siarhei Kharytonchyk ◽  
Alexis Waller ◽  
Ugonna Mbaekwe ◽  
Sapna Basappa ◽  
...  
Keyword(s):  
Binding Sites ◽  
Virus Assembly ◽  
Rna Packaging ◽  
Packaging Signal ◽  
Nmr Studies ◽  
Recognition Element ◽  
High Affinity ◽  
Molecule Inhibitor ◽  
Encapsidation Signal ◽  
Hiv 1

Selective packaging of the HIV-1 genome during virus assembly is mediated by interactions between the dimeric 5ʹ-leader of the unspliced viral RNA and the nucleocapsid (NC) domains of a small number of assembling viral Gag polyproteins. Here, we show that the dimeric 5′-leader contains more than two dozen NC binding sites with affinities ranging from 40 nM to 1.4 μM, and that all high-affinity sites (Kd≲ 400 nM) reside within a ∼150-nt region of the leader sufficient to promote RNA packaging (core encapsidation signal, ΨCES). The four initial binding sites with highest affinity reside near two symmetrically equivalent three-way junction structures. Unlike the other high-affinity sites, which bind NC with exothermic energetics, binding to these sites occurs endothermically due to concomitant unwinding of a weakly base-paired [UUUU]:[GGAG] helical element. Mutations that stabilize base pairing within this element eliminate NC binding to this site and severely impair RNA packaging into virus-like particles. NMR studies reveal that a recently discovered small-molecule inhibitor of HIV-1 RNA packaging that appears to function by stabilizing the structure of the leader binds directly to the [UUUU]:[GGAG] helix. Our findings suggest a sequential NC binding mechanism for Gag-genome assembly and identify a potential RNA Achilles’ heel to which HIV therapeutics may be targeted.

scholarly journals STD and TRNOESY NMR studies for the epitope mapping of the phosphorylation motif of the oncogenic protein β-catenin recognized by a selective monoclonal antibody

FEBS Letters ◽  
2006 ◽  
Vol 580 (22) ◽  
pp. 5411-5422 ◽  
Author(s):  
Simon Megy ◽  
Gildas Bertho ◽  
Josyane Gharbi-Benarous ◽  
Françoise Baleux ◽  
Richard Benarous ◽  
...  
Keyword(s):  
Monoclonal Antibody ◽  
Epitope Mapping ◽  
Nmr Studies ◽  
Phosphorylation Motif ◽  
Oncogenic Protein

scholarly journals Skp1 Dimerization Conceals its F-box Protein Binding Site

2019 ◽  
Author(s):  
Hyun W. Kim ◽  
Alexander Eletsky ◽  
Karen J. Gonzalez ◽  
Hanke van der Wel ◽  
Eva-Maria Strauch ◽  
...  
Keyword(s):  
Solution Structure ◽  
Hydrophobic Surface ◽  
Solution Nmr ◽  
Nmr Studies ◽  
Dimer Interface ◽  
High Resolution Nmr ◽  
Specific Proteins ◽  
The Difference ◽  
F Box Protein

ABSTRACTSkp1 is an adapter that links F-box proteins to cullin-1 in the Skp1/cullin-1/F-box (SCF) protein family of E3 ubiquitin ligases that targets specific proteins for polyubiquitination and subsequent protein degradation. Skp1 from the amoebozoan Dictyostelium forms a stable homodimer in vitro with a Kd of 2.5 µM as determined by sedimentation velocity studies, yet is monomeric in crystal complexes with F-box proteins. To investigate the molecular basis for the difference, we determined the solution NMR structure of a doubly truncated Skp1 homodimer (Skp1ΔΔ). The solution structure of Skp1ΔΔ dimer reveals a 2-fold symmetry with an interface that buries ∼750 Å2 of predominantly hydrophobic surface. The dimer interface overlaps with subsite-1 of the F-box interaction area, explaining why only the Skp1 monomer binds F-box proteins (FBPs). To confirm the model, Rosetta was used to predict amino acid substitutions that might disrupt the dimer interface, and the F97E substitution was chosen to potentially minimize interference with F-box interactions. A nearly full-length version of Skp1 with this substitution (Skp1ΔF97E) behaved as a stable monomer at concentrations up to 500 µM and actively bound a model FBP, mammalian Fbs1, which suggests that the dimeric state is not required for Skp1 to carry out a basic biochemical function. Finally, Skp1ΔF97E is expected to serve as a monomer model for high-resolution NMR studies previously hindered by dimerization.

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