Journal: Cell reports

Article Title: Localized phosphorylation of a kinesin-1 adaptor by a capsid-associated kinase regulates HIV-1 motility and uncoating

doi: 10.1016/j.celrep.2017.08.076

Figure Lengend Snippet: (A) GFP-tagged MARK2 (GFP-MARK2) expression in 293T cells results in phosphorylation of FEZ1-Flag at S58, which is not detected with a FEZ1-S58A-Flag mutant. (B) Time course infection demonstrating HIV-1 does not induce phosphorylation of FEZ1 in HIV-1-VSV infected 293T cells at indicated h.p.i. (C) Similar to FEZ1-Flag, GFP-MARK2, but not control GAPDH-HA, binds to in vitro assembled HIV-1 CA-NC complexes. (D) GFP-MARK2, but not GFP control, increases FEZ1 phosphorylation at S58 in 293T cells co-expressing GFP-MARK2 and FEZ1-Flag. Results in A–D are representative of three or more independent experiments.

Article Snippet: For transient expression of control GFP or GFP-tagged MARK2, 293T cells were transfected with 1.2 μg of pEGFP-C1 or a C′ terminally GFP-tagged MARK2 (Addgene plasmid # 66706, deposited by Gary Banker) or a N′ terminally GFP-tagged MARK2 (GFP-MARK2) expressing construct (pEGFP-EMK C1, kindly provided by Andrew S. Shaw)( Lin et al., 2009 ) in a 12-well plate or with 5–10 μg of each of the constructs in a 100 mm dish using polyethylenimine.

Techniques: Expressing, Mutagenesis, Infection, In Vitro

Journal: Cell reports

Article Title: Localized phosphorylation of a kinesin-1 adaptor by a capsid-associated kinase regulates HIV-1 motility and uncoating

doi: 10.1016/j.celrep.2017.08.076

Figure Lengend Snippet: (A–B) Transient or stable expression of MARK2-GFP, but not control GFP, increases infection by HIV-1-VSV-Luc in 293T cells (A) or HIV-1-WT-Luc in Jurkat cells (B) at different virus dilutions. (C) RNAi-mediated FEZ1 depletion abrogates the ability of MARK2-GFP to enhance HIV-1-VSV-Luc infection. Data in A–C, upper panels, are shown as mean ± SD, n= 3.

Article Snippet: For transient expression of control GFP or GFP-tagged MARK2, 293T cells were transfected with 1.2 μg of pEGFP-C1 or a C′ terminally GFP-tagged MARK2 (Addgene plasmid # 66706, deposited by Gary Banker) or a N′ terminally GFP-tagged MARK2 (GFP-MARK2) expressing construct (pEGFP-EMK C1, kindly provided by Andrew S. Shaw)( Lin et al., 2009 ) in a 12-well plate or with 5–10 μg of each of the constructs in a 100 mm dish using polyethylenimine.

Techniques: Expressing, Infection