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Many of these conclusions could be drawn from a recently published electron microscopic reconstruction of an RNAP Spt4/5 complex (Klein et al, 2011) and are confirmed here and extended based on high-resolution data.
Cryo-electron microscopic reconstruction of eIF3 has revealed that its subunits are organized in an anthropomorphic shape with five appendages and which shows surface complementarity to the platform of the 40S ribosomal subunit [ 10].
Electron microscopic reconstruction of the amphids of the free-living nematode C. elegans and several parasitic nematodes such as H. contortus and Ancylostoma caninum, have shown that the sensory neuroanatomy is remarkably similar between the different species [ 1, 6, 7, 10, 11].
After our work had been completed, an electron microscopic reconstruction of an RNAP Spt4/5 complex was reported that provided a medium-resolution view of the Spt4/5-containing RNAP elongation complex and resulted in similar overall conclusions (Klein et al, 2011).
Furthermore, electron microscopic reconstruction of the Pyrococcus furiosus clamp loader demonstrates that archaeal clamp loaders also have the A' domain and shows electron density that reaches across the interface and touches the clamp, like the T4 loader [ 60, 62] but unlike the E. coli A subunit, which binds only one protomer of the clamp and contains no A' domain [ 32, 114].
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The RNCs generated with this approach have been successfully used to obtain 3D cryo-electron microscopic reconstructions of complexes with the signal recognition particle and the translocon.
The CCDB was formally launched in 2002 as an on-line repository of high-resolution 3D light and electron microscopic reconstructions of cells and subcellular structures.
Both electron microscopic reconstructions of Myo1A decorating microfilaments and biophysical assays have demonstrated that this protein is a fully functional plus-end directed myosin [5], [29] and that it binds actin in a manner indistinguishable from that of conventional, class-II myosins [30], [31].
An electron cryo-microscopic reconstruction of these complexes reveals their architecture in new detail, and shows that the complex is constructed from four SecY/Sec61 heterotrimers and two TRAP complexes.
Sampled synapses are mapped onto previous light-microscopic reconstructions of the same neurons (Glaser and Glaser 1990; Ascoli 2006) to examine the distribution and density of inputs within different somato-dendritic compartments, thus defining key features of the neuron's afferent 'synaptome' (DeFelipe 2010).
After light microscopic reconstruction, stained cells were serially sectioned into 90 nm thickness using an ultramicrotome (Reichert Ultracut S).
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