Biomedical subjects

Harald Dürr

Publications and source records attributed to Harald Dürr.

4 recordsLinked to original sources

PubMed2006

Snf2 family ATPases and DExx box helicases: differences and unifying concepts from high-resolution crystal structures.

Proteins with sequence similarity to the yeast Snf2 protein form a large family of ATPases that act to alter the structure of a diverse range of DNA-protein structures including chromatin. Snf2 family enzymes are related in sequence to DExx box helicases, yet they do not possess helicase activity. Recent biochemical and structural studies suggest that the mechanism by which these enzymes act involves ATP-dependent translocation on DNA. Crystal structures suggest that these enzymes travel along the minor groove, a process that can generate the torque or energy in remodelling processes. We review the recent structural and biochemical findings which suggest a common mechanistic basis underlies the action of many of both Snf2 family and DExx box helicases.

Adenosine Triphosphatases↗

PubMed2006

Structure-function analysis of SWI2/SNF2 enzymes.

Biochemical and structural progress over the last years has revealed that SWI2/SNF2 family chromatin remodeling or DNA repair enzymes are molecular motors that transport duplex DNA along a helicase-like domain using ATP-hydrolysis. The screw motion of DNA along the active site probably generates the force to disrupt chromatin or other protein:DNA complexes. In this chapter, we describe biochemical and structural approaches to study the molecular mechanism of SWI2/SNF2 enzymes. In particular, we describe assays to monitor DNA dependent ATPase activity, translocation on duplex DNA, and DNA distortion activity. We also describe recent progress in the crystallization and structure determination of SWI2/SNF2 enzymes in complex with duplex DNA.

Adenosine Triphosphatases↗

PubMed2005

X-ray structures of the Sulfolobus solfataricus SWI2/SNF2 ATPase core and its complex with DNA.

SWI2/SNF2 ATPases remodel chromatin or other DNA:protein complexes by a poorly understood mechanism that involves ATP-dependent DNA translocation and generation of superhelical torsion. Crystal structures of a dsDNA-translocating SWI2/SNF2 ATPase core from Sulfolobus solfataricus reveal two helical SWI2/SNF2 specific subdomains, fused to a DExx box helicase-related ATPase core. Fully base paired duplex DNA binds along a central cleft via both minor groove strands, indicating that SWI2/SNF2 ATPases travel along the dsDNA minor groove without strand separation. A structural switch, linking DNA binding and the active site DExx motif, may account for the stimulation of ATPase activity by dsDNA. Our results suggest that torque in remodeling processes is generated by an ATP-driven screw motion of DNA along the active site cleft. The structures also redefine SWI2/SNF2 functional motifs and uncover unexpected structural correlation of mutations in Cockayne and X-linked mental retardation syndromes.

Adenosine Triphosphatases↗

PubMed2005

Chromophore exchange in the LOV2 domain of the plant photoreceptor phototropin1 from oat.

Phototropins are a family of plant photoreceptors mediating blue light responses such as phototropism, leaf expansion, chloroplast relocation, and stomatal opening. Characteristic for phototropins are two LOV domains which, when expressed in heterologous systems, each carry a single flavin mononucleotide (FMN) chromophore. Here we describe removal of FMN from the LOV2 domain of Avena sativa using a hydrophobic matrix and successful incorporation of flavin adenine dinucleotide (FAD), riboflavin, and 5'-malonyl-riboflavin into the resulting apoprotein; 5-deaza-FMN was not incorporated under the applied conditions. The chromoproteins reconstituted with the various flavins showed absorption spectra and photocycle almost identical to those of the native LOV2 domain and that reconstituted with FMN except for the kinetics: LOV2-riboflavin and LOV2-5'-malonyl-riboflavin showed more rapid regeneration in the dark. LOV2-FAD can be hydrolyzed to LOV2-FMN with phosphodiesterase, indicating that the adenosine part extrudes from the protein. Together with the data from the X-ray structure (Crosson, S., and Moffat, K. (2001) Proc. Natl. Acad. Sci. U.S.A. 98, 2995-3000), the results allow us to decide which of the chromophore-protein interactions are essential for the reconstitution process.

Avena↗