7MI8

Signal subtracted reconstruction of AAA5 and AAA6 domains of dynein in the presence of a pyrazolo-pyrimidinone-based compound, Model 5

Summary for 7MI8

• Entry DOI: 10.2210/pdb7mi8/pdb
• EMDB information: 23842
• Descriptor: Fusion protein of Dynein and Endolysin (1 entity in total)
• Functional Keywords: aaa atpase, atpase inhibitor, motor protein
• Biological source: Saccharomyces cerevisiae (Baker's yeast); More
• Total number of polymer chains: 1
• Total formula weight: 304889.88

Authors

Santarossa, C.C.,Coudray, N.,Urnavicius, L.,Ekiert, D.C.,Bhabha, G.,Kapoor, T.M. (deposition date: 2021-04-16, release date: 2021-05-26, Last modification date: 2024-05-29)

Primary citation

Santarossa, C.C.,Mickolajczyk, K.J.,Steinman, J.B.,Urnavicius, L.,Chen, N.,Hirata, Y.,Fukase, Y.,Coudray, N.,Ekiert, D.C.,Bhabha, G.,Kapoor, T.M.
Targeting allostery in the Dynein motor domain with small molecule inhibitors.
Cell Chem Biol, 28:1460-1473.e15, 2021

PubMed Abstract: Cytoplasmic dyneins are AAA (ATPase associated with diverse cellular activities) motor proteins responsible for microtubule minus-end-directed intracellular transport. Dynein's unusually large size, four distinct nucleotide-binding sites, and conformational dynamics pose challenges for the design of potent and selective chemical inhibitors. Here we use structural approaches to develop a model for the inhibition of a well-characterized S. cerevisiae dynein construct by pyrazolo-pyrimidinone-based compounds. These data, along with functional assays of dynein motility and mutagenesis studies, suggest that the compounds inhibit dynein by engaging the regulatory ATPase sites in the AAA3 and AAA4 domains, and not by interacting with dynein's main catalytic site in the AAA1 domain. A double Walker B mutation of the AAA3 and AAA4 sites substantially reduces enzyme activity, suggesting that targeting these regulatory domains is sufficient to inhibit dynein. Our findings reveal how chemical inhibitors can be designed to disrupt allosteric communication across dynein's AAA domains.

PubMed: 34015309
DOI: 10.1016/j.chembiol.2021.04.024

Experimental method

ELECTRON MICROSCOPY (3.7 Å)