Molecular mechanism of force generation by the cytoplasmic dynein motor protein complex at atomic resolution
Final Report Abstract
Within this project we have identified, assembled and annotated several thousand dynein, dynactin and tubulin sequences. Based on the sequence data, the evolution of the dynactin complex in eukaryotes has been reconstructed. The dynein data helped in characterising domain borders, based on sequence homology and based on the identification of many split dynein heavy chain genes with so far unknown split sites. We were able to express several dozen fragments of the DHC genes of Dictyostelium, human (both DHC1 and DHC7), and yeast (S.cerevisiae). Two of these fragments could be crystallized and the crystallization conditions optimized so that nicelooking, large crystals can reproducibly be obtained. Unfortunately, the diffraction quality could in general not be improved to better than 6-7 Å, although one crystal even diffracted to better than 3.5 Å. We obtained high-resolution structures of parts of the dynactin complex, the barbedend capping protein complex Cap32/34 and the fragment of the p150 (dynactin-1) subunit, that binds to the dynein intermediate chain.
Publications
- (2011) Cross‐ species protein sequence and gene structure prediction with fine‐tuned Webscipio 2.0 and Scipio. BMC Research Notes 4, 265, 1‐20
Hatje, K., Keller, O., Hammesfahr, B., Pillmann, H., Waack, S. & Kollmar, M.
- (2011) diArk 2.0 provides detailed analyses of the ever increasing eukaryotic genome sequencing data. BMC Research Notes 4, 338, 1‐12
Hammesfahr, B., Odronitz, F., Hellkamp, M. & Kollmar, M.
- (2011) Predicting mutually exclusive spliced exons based on exon length, splice site and reading frame conservation, and exon sequence homology. BMC Bioinformatics 12, 270, 1‐16
Pillmann, H., Hatje, K., Odronitz, F., Hammesfahr, B. & Kollmar, M.
- (2012) Conservation and divergence between cytoplasmic and muscle‐specific actin capping proteins: insights from the crystal structure of cytoplasmic Cap32/34 from Dictyostelium discoideum. BMC Structural Biology 12, 12, 1‐ 15
Eckert, C., Goretzki, A., Faberova, M. & Kollmar, M.
(See online at https://doi.org/10.1186/1472-6807-12-12) - (2012) Evolution of the eukaryotic dynactin complex, the activator of cytoplasmic dynein. BMC Evolutionary Biology 12, 95, 1‐21
Hammesfahr, B. & Kollmar, M.
(See online at https://doi.org/10.1186/1471-2148-12-95) - (2014) Six subgroups and extensive recent duplications characterize the evolution of the eukaryotic tubulin protein family. Genome Biology and Evolution 6, 2274‐2288
Findeisen, P., Mühlhausen, S., Dempewolf, S., Hertzog, J., Zietlow, A., Carlomagno, T. & Kollmar, M.
(See online at https://doi.org/10.1093/gbe/evu187)
