Regulation of Human Hsp70 by its Nucleotide Exchange Factors (NEFs).

Abstract

Heat shock protein 70 (Hsp70) is an abundant and ubiquitous molecular chaperone that is responsible for maintenance of the human proteome. Accordingly, Hsp70 has become an attractive drug target for neurodegenerative and hyperproliferative disorders; however it is difficult to imagine strategies for inhibiting its pathobiology without impacting its essential roles. Fortunately, Hsp70 does not work alone, and instead employs a large network of co-chaperone proteins, which can tune Hsp70 activity and influence disease state. These co-chaperone proteins provide potential handles for targeting Hsp70 without disrupting overall proteostasis.

One such class of co-chaperones proteins known as the Nucleotide Exchange Factors (NEFs), are a particular appealing target. NEFs bind Hsp70 and help to facilitate the exchange of ADP for ATP. The biochemistry of the NEF family of co-chaperones has classically been investigated using the prokaryotic NEF, GrpE, as a model. However, the eukaryotic cytosol does not contain a GrpE homolog. Rather, there are three main sub-classes of human NEFs: Hsp110, HspBP1, and the BAG proteins, all of which are structurally distinct with little sequence homology. Consistent with their diverse structures, they also differ in their mode of binding to Hsp70 and their roles in guiding Hsp70 biology. For example, BAG2 is associated with proteasomal degradation of the Hsp70 substrate, tau, while BAG1-Hsp70 is linked to increased tau stability. These observations suggest that the formation of specific NEF-Hsp70 complexes may help decide the fate of Hsp70-bound substrates. Additionally, these findings illustrate that differential disruption of specific Hsp70-NEF contacts might be beneficial in disease.

In this thesis work I have systematically characterized the human Hsp70 NEFs, including how they interact with Hsp70, how the influence Hsp70 biochemistry and how they can bridge Hsp70 with other classes of chaperone proteins. I have used high throughput screening methods to search for chemical matter that can modulate Hsp70-NEF interactions, and we have shown that inhibitors of Hsp70-NEF interactions can be beneficial for treating disease. This thesis work has significantly advanced our knowledge of human Hsp70 regulation, and has provided groundwork for future studies on other Hsp70 co-chaperones and proteostasis components.