DEAD-box proteins catalyze the ATP-dependent unwinding of RNA duplexes. The common unit of these enzymes is a helicase core of two flexibly linked RecA domains. ATP binding and phosphate release control opening and closing of the cleft in the helicase core. This movement coordinates RNA-binding and ATPase activity and is thus central to the function of DEAD-box helicases. In most DEAD box proteins, the helicase core is flanked by ancillary N-and C-terminal domains. Here, we describe single molecule fluorescence resonance energy transfer (smFRET) approaches to directly monitor conformational changes associated with opening and closing of the helicase core. We further outline smFRET strategies to determine the orientation of flanking N- and C-terminal domains of DEAD-box helicases and to assess the effects of regulatory proteins on DEAD-box helicase conformation.
机构:
Stanford Univ, Howard Hughes Med Inst, Stanford, CA 94305 USA
Stanford Univ, Dept Mol & Cellular Physiol, Stanford, CA 94305 USA
Stanford Univ, Dept Neurol & Neurol Sci, Stanford, CA 94305 USA
Stanford Univ, Dept Biol Struct, Stanford, CA 94305 USA
Stanford Univ, Dept Photon Sci, Stanford, CA 94305 USAStanford Univ, Howard Hughes Med Inst, Stanford, CA 94305 USA
Brunger, Axel T.
Strop, Pavel
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Stanford Univ, Howard Hughes Med Inst, Stanford, CA 94305 USA
Stanford Univ, Dept Mol & Cellular Physiol, Stanford, CA 94305 USA
Stanford Univ, Dept Neurol & Neurol Sci, Stanford, CA 94305 USA
Stanford Univ, Dept Biol Struct, Stanford, CA 94305 USA
Stanford Univ, Dept Photon Sci, Stanford, CA 94305 USAStanford Univ, Howard Hughes Med Inst, Stanford, CA 94305 USA
Strop, Pavel
Vrljic, Marija
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Stanford Univ, Howard Hughes Med Inst, Stanford, CA 94305 USA
Stanford Univ, Dept Mol & Cellular Physiol, Stanford, CA 94305 USA
Stanford Univ, Dept Neurol & Neurol Sci, Stanford, CA 94305 USA
Stanford Univ, Dept Biol Struct, Stanford, CA 94305 USA
Stanford Univ, Dept Photon Sci, Stanford, CA 94305 USAStanford Univ, Howard Hughes Med Inst, Stanford, CA 94305 USA
Vrljic, Marija
Chu, Steven
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Univ Calif Berkeley, Dept Phys, Berkeley, CA 94720 USA
Univ Calif Berkeley, Dept Mol & Cell Biol, Berkeley, CA 94720 USA
Lawrence Berkeley Lab, Berkeley, CA 94720 USAStanford Univ, Howard Hughes Med Inst, Stanford, CA 94305 USA
Chu, Steven
Weninger, Keith R.
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N Carolina State Univ, Dept Phys, Raleigh, NC 27695 USAStanford Univ, Howard Hughes Med Inst, Stanford, CA 94305 USA
机构:
Stanford Univ, Howard Hughes Med Inst, Stanford, CA 94305 USA
Stanford Univ, Dept Mol & Cellular Physiol, Stanford, CA 94305 USA
Stanford Univ, Dept Neurol & Neurol Sci, Stanford, CA 94305 USA
Stanford Univ, Dept Biol Struct, Stanford, CA 94305 USA
Stanford Univ, Dept Photon Sci, Stanford, CA 94305 USAStanford Univ, Howard Hughes Med Inst, Stanford, CA 94305 USA
Brunger, Axel T.
Strop, Pavel
论文数: 0引用数: 0
h-index: 0
机构:
Stanford Univ, Howard Hughes Med Inst, Stanford, CA 94305 USA
Stanford Univ, Dept Mol & Cellular Physiol, Stanford, CA 94305 USA
Stanford Univ, Dept Neurol & Neurol Sci, Stanford, CA 94305 USA
Stanford Univ, Dept Biol Struct, Stanford, CA 94305 USA
Stanford Univ, Dept Photon Sci, Stanford, CA 94305 USAStanford Univ, Howard Hughes Med Inst, Stanford, CA 94305 USA
Strop, Pavel
Vrljic, Marija
论文数: 0引用数: 0
h-index: 0
机构:
Stanford Univ, Howard Hughes Med Inst, Stanford, CA 94305 USA
Stanford Univ, Dept Mol & Cellular Physiol, Stanford, CA 94305 USA
Stanford Univ, Dept Neurol & Neurol Sci, Stanford, CA 94305 USA
Stanford Univ, Dept Biol Struct, Stanford, CA 94305 USA
Stanford Univ, Dept Photon Sci, Stanford, CA 94305 USAStanford Univ, Howard Hughes Med Inst, Stanford, CA 94305 USA
Vrljic, Marija
Chu, Steven
论文数: 0引用数: 0
h-index: 0
机构:
Univ Calif Berkeley, Dept Phys, Berkeley, CA 94720 USA
Univ Calif Berkeley, Dept Mol & Cell Biol, Berkeley, CA 94720 USA
Lawrence Berkeley Lab, Berkeley, CA 94720 USAStanford Univ, Howard Hughes Med Inst, Stanford, CA 94305 USA
Chu, Steven
Weninger, Keith R.
论文数: 0引用数: 0
h-index: 0
机构:
N Carolina State Univ, Dept Phys, Raleigh, NC 27695 USAStanford Univ, Howard Hughes Med Inst, Stanford, CA 94305 USA