Two-dimensional averaged images of the dynactin complex revealed by single particle analysis.
The dynactin complex interacts with dynein and numerous other proteins to provide for a wide range of subcellular transport functions. A detailed understanding of the structure and subunit organization of dynactin should yield new insights into its function. In the present study, we used single particle analysis to obtain a two-dimensional averaged image of dynactin isolated from chick embryo brains and visualized by negative stain electron microscopy (EM). Each individual image, consisting of the shoulder/sidearm and the rod, closely resembled the previously published quick-freeze deep-etch rotary-shadow electron micrographs. However, the averaged image revealed novel structural features that may have functional significance. The bulky shoulder complex has a triangular shape and is 13 nm wide and 8 nm high. The rod, with an overall length of 40 nm, consists of clearly defined lobes that are apparently grouped into three parts, the pointed-end complex, the middle segment, and the extra lobes at the barbed end. The pointed-end complex reveals the characteristic protrusions and clefts that were previously observed only in the isolated pointed-end complex. In the middle segment, the seven lobes are fitted to the helical symmetry of F-actin. A narrow but prominent gap separates the previously unidentified extra three lobes at the barbed end from the middle segment. The averaged image we obtained contrasts dramatically with the simple Arp1 polymer that was previously reported by single particle analysis of bovine brain dynactin. These apparent structural differences are probably due to the greater stability and integrity of the chick embryo brain dynactin preparation. We propose a new structural model for dynactin, based on our observations.
Imai
Hiroshi
H
ERATO Actin Filament Dynamics Project, Japan Science and Technology Agency c/o RIKEN, Sayo, Hyogo 679-5148, Japan.
Narita
Akihiro
A
Schroer
Trina A
TA
Maéda
Yuichiro
Y
D008841:Actin Cytoskeleton / Q000737:chemistry; D000199:Actins / Q000737:chemistry; D000818:Animals; D001923:Brain Chemistry; D051837:COUP Transcription Factor II / Q000737:chemistry; D002642:Chick Embryo; D000072159:Dynactin Complex; D007091:Image Processing, Computer-Assisted / Q000379:methods*; D008854:Microscopy, Electron / Q000379:methods*; D008869:Microtubule-Associated Proteins / Q000737:chemistry*; D008958:Models, Molecular; D011487:Protein Conformation
D000199:Actins; D051837:COUP Transcription Factor II; D000072159:Dynactin Complex; D008869:Microtubule-Associated Proteins; C491475:actin-related protein 1, rat; C095569:NR2F2 protein, human