Journal: Sci Adv / Year: 2017 Title: CryoEM structure of MxB reveals a novel oligomerization interface critical for HIV restriction. Authors: Frances J D Alvarez / Shaoda He / Juan R Perilla / Sooin Jang / Klaus Schulten / Alan N Engelman / Sjors H W Scheres / Peijun Zhang / Abstract: Human dynamin-like, interferon-induced myxovirus resistance 2 (Mx2 or MxB) is a potent HIV-1 inhibitor. Antiviral activity requires both the amino-terminal region of MxB and protein oligomerization, ...Human dynamin-like, interferon-induced myxovirus resistance 2 (Mx2 or MxB) is a potent HIV-1 inhibitor. Antiviral activity requires both the amino-terminal region of MxB and protein oligomerization, each of which has eluded structural determination due to difficulties in protein preparation. We report that maltose binding protein-fused, full-length wild-type MxB purifies as oligomers and further self-assembles into helical arrays in physiological salt. Guanosine triphosphate (GTP), but not guanosine diphosphate, binding results in array disassembly, whereas subsequent GTP hydrolysis allows its reformation. Using cryo-electron microscopy (cryoEM), we determined the MxB assembly structure at 4.6 Å resolution, representing the first near-atomic resolution structure in the mammalian dynamin superfamily. The structure revealed previously described and novel MxB assembly interfaces. Mutational analyses demonstrated a critical role for one of the novel interfaces in HIV-1 restriction.
0: Interferon-induced GTP-binding protein Mx2 1: Interferon-induced GTP-binding protein Mx2 2: Interferon-induced GTP-binding protein Mx2 3: Interferon-induced GTP-binding protein Mx2 4: Interferon-induced GTP-binding protein Mx2 5: Interferon-induced GTP-binding protein Mx2 6: Interferon-induced GTP-binding protein Mx2 7: Interferon-induced GTP-binding protein Mx2 8: Interferon-induced GTP-binding protein Mx2 9: Interferon-induced GTP-binding protein Mx2 a: Interferon-induced GTP-binding protein Mx2 b: Interferon-induced GTP-binding protein Mx2 c: Interferon-induced GTP-binding protein Mx2 d: Interferon-induced GTP-binding protein Mx2 e: Interferon-induced GTP-binding protein Mx2 f: Interferon-induced GTP-binding protein Mx2 g: Interferon-induced GTP-binding protein Mx2 h: Interferon-induced GTP-binding protein Mx2 i: Interferon-induced GTP-binding protein Mx2 j: Interferon-induced GTP-binding protein Mx2 k: Interferon-induced GTP-binding protein Mx2 l: Interferon-induced GTP-binding protein Mx2 m: Interferon-induced GTP-binding protein Mx2 n: Interferon-induced GTP-binding protein Mx2 o: Interferon-induced GTP-binding protein Mx2 p: Interferon-induced GTP-binding protein Mx2 q: Interferon-induced GTP-binding protein Mx2 r: Interferon-induced GTP-binding protein Mx2 s: Interferon-induced GTP-binding protein Mx2 t: Interferon-induced GTP-binding protein Mx2 u: Interferon-induced GTP-binding protein Mx2 v: Interferon-induced GTP-binding protein Mx2 w: Interferon-induced GTP-binding protein Mx2 x: Interferon-induced GTP-binding protein Mx2 y: Interferon-induced GTP-binding protein Mx2 z: Interferon-induced GTP-binding protein Mx2 A: Interferon-induced GTP-binding protein Mx2 B: Interferon-induced GTP-binding protein Mx2 C: Interferon-induced GTP-binding protein Mx2 D: Interferon-induced GTP-binding protein Mx2 E: Interferon-induced GTP-binding protein Mx2 F: Interferon-induced GTP-binding protein Mx2 G: Interferon-induced GTP-binding protein Mx2 H: Interferon-induced GTP-binding protein Mx2 I: Interferon-induced GTP-binding protein Mx2 J: Interferon-induced GTP-binding protein Mx2 K: Interferon-induced GTP-binding protein Mx2 L: Interferon-induced GTP-binding protein Mx2 M: Interferon-induced GTP-binding protein Mx2 N: Interferon-induced GTP-binding protein Mx2 O: Interferon-induced GTP-binding protein Mx2 P: Interferon-induced GTP-binding protein Mx2 Q: Interferon-induced GTP-binding protein Mx2 R: Interferon-induced GTP-binding protein Mx2 S: Interferon-induced GTP-binding protein Mx2 T: Interferon-induced GTP-binding protein Mx2 U: Interferon-induced GTP-binding protein Mx2 V: Interferon-induced GTP-binding protein Mx2 W: Interferon-induced GTP-binding protein Mx2 X: Interferon-induced GTP-binding protein Mx2 Y: Interferon-induced GTP-binding protein Mx2 Z: Interferon-induced GTP-binding protein Mx2
Model: Tecnai Polara / Image courtesy: FEI Company
Microscopy
Model: FEI POLARA 300
Electron gun
Electron source: FIELD EMISSION GUN / Accelerating voltage: 300 kV / Illumination mode: FLOOD BEAM
Electron lens
Mode: BRIGHT FIELD / Nominal magnification: 93000 X / Nominal defocus max: 3500 nm / Nominal defocus min: 1000 nm / Cs: 2.2 mm / C2 aperture diameter: 100 µm
Specimen holder
Cryogen: NITROGEN
Image recording
Average exposure time: 1.6 sec. / Electron dose: 40 e/Å2 / Detector mode: INTEGRATING / Film or detector model: FEI FALCON II (4k x 4k) / Num. of grids imaged: 1 / Num. of real images: 1123
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