Journal: The Journal of Biological Chemistry

Article Title: Cyclin F/FBXO1 Interacts with HIV-1 Viral Infectivity Factor (Vif) and Restricts Progeny Virion Infectivity by Ubiquitination and Proteasomal Degradation of Vif Protein through SCF cyclin F E3 Ligase Machinery *

doi: 10.1074/jbc.M116.765842

Figure Lengend Snippet: Cyclin F physically interacts with Vif during HIV-1 infection in T cells. A , co-immunoprecipitation and reverse co-immunoprecipitation showing interaction of cyclin F and HIV-1 Vif in transfected HEK293T cells. B , cyclin F and Vif interacts endogenously during HIV-1 infection in 0.5 m.o.i. infected CEM-GFP (72 hpi) cells. Skp1 is also co-immunoprecipitated with cyclin F and Vif pulldown. C , cyclin F and Vif is present in the nucleus ( NE ) as well as the cytoplasm ( CE ) during infection. D , co-localization of cyclin F and Vif in both nuclear and cytoplasmic compartments in HIV-1-infected TZM-bl cells observed at 24 h post-infection. All panels are representative of at least three independent experiments. IP , immunoprecipitation; IB , immunoblot.

Article Snippet: The Skp1 antibody (rabbit, catalog No. 100-401-A08, lot 15426) was procured from Rockland Immunochemicals.

Techniques: Infection, Immunoprecipitation, Transfection, Western Blot

Journal: The Journal of Biological Chemistry

Article Title: Cyclin F/FBXO1 Interacts with HIV-1 Viral Infectivity Factor (Vif) and Restricts Progeny Virion Infectivity by Ubiquitination and Proteasomal Degradation of Vif Protein through SCF cyclin F E3 Ligase Machinery *

doi: 10.1074/jbc.M116.765842

Figure Lengend Snippet: Cyclin F binds to Vif through the CY motif in the C-terminal region of HIV-1 NL4-3 Vif. A , molecular docking of modeled cyclin domain region (including cyclin C domain) of cyclin F with available HIV-1 Vif structure. Amino acid residues in cyclin F and Vif involved in the interaction as predicted by PatchDock Server are shown in the zoomed image below. B , depiction of CY mutant generation of Vif ( upper panel ). Cyclin F does not co-immunoprecipitate CY mutant Vif ( lower panel ). Cyclin F-Skp1 interaction is found to be intact, but cyclin F-Vif interaction is lost when CY mutant of Vif is co-transfected with cyclin F in transfected 293T cells. All panels represent data from at least two or more independent experiments. IP , immunoprecipitate; IB , immunoblot. C , CY mutant Vif is not down-regulated by cyclin F in transfected 293T cells ( n = 2). Data represent mean ± S.E.

Article Snippet: The Skp1 antibody (rabbit, catalog No. 100-401-A08, lot 15426) was procured from Rockland Immunochemicals.

Techniques: Mutagenesis, Transfection, Western Blot

Journal: STAR Protocols

Article Title: Protocol for analyzing intact mRNA poly(A) tail length using nanopore direct RNA sequencing

doi: 10.1016/j.xpro.2023.102340

Figure Lengend Snippet: Preparation of GST-eIF4E K119A , input RNA and cap-purified RNA (A) Small fraction of samples in the indicated steps are resolved by SDS-PAGE and proteins were visualized with CBB. IPTG(-): before induction, IPTG(+): after induction, Sonicated: sonicated sample before centrifugation, Cleared sup: sonicated sample after centrifugation, Flow-through: proteins unbound to glutathione 4B Sepharose, Eluted1/2/3: proteins eluted using glutathione, Beads: proteins remained on the beads after three consecutive elution. (B) EtBr staining showing efficient removal of small RNAs by two consecutive LiCl precipitation. ppt: precipitated, sup: unprecipitated fraction was ethanol-precipitated and then analyzed. (C) CBB staining of samples after glutathione/3C protease elution of capped RNAs. (D) RNA samples in the indicated steps are resolved by agarose gel electrophoresis and stained with SYBR Gold. Recombinant GST protein was used in the control pull down experiments.

Article Snippet: pGEX6P-1/eIF4E K119A , Addgene , Addgene: 199440.

Techniques: Purification, SDS Page, Sonication, Centrifugation, Staining, Agarose Gel Electrophoresis, Recombinant, Control

Journal: STAR Protocols

Article Title: Protocol for analyzing intact mRNA poly(A) tail length using nanopore direct RNA sequencing

doi: 10.1016/j.xpro.2023.102340

Figure Lengend Snippet:

Article Snippet: pGEX6P-1/eIF4E K119A , Addgene , Addgene: 199440.

Techniques: Virus, Recombinant, Protease Inhibitor, RNA Sequencing, Software