Understanding the Hsp70 chaperone code
Hsp70 is a universally conserved molecular chaperone that performs a wide variety of functions in the cell including folding of both newly synthesized and denatured protein “clients”. Although transcriptional regulation of Hsp70 has been highly studied, little is known about post-translational modifications on Hsp70 (known as the “Chaperone Code”) and their effects on in vivo function. We are studying the conditions that alter the PTMs on Hsp70 in yeast and human cells and the effect they have on in vivo function. For more information please see our latest review on Hsp70 PTMs here
Understanding the role of DNAJA1 in cancer
Our previous studies uncovered a role for Hsp70, Hsp90 and DNAJA1 in the stability of Ribonucleotide reductase (RNR), an important enzyme required for repairing DNA. Following up from this, we performed a chemogenomic screen to identify the role of DNAJA1 in anticancer drug resistance. A substantial number of drugs become far more potent upon loss of DNAJA1. We validate several of these using the DNAJA1 inhibitor 116-9e on both prostate cancer cell culture and spheroids models. We are now hoping to study these synergistic combinations in mice models.
Why do cells express so many highly-related and seemingly-related chaperones? Our research suggests that while they have overlapping roles, there are clear functional differences between them. We are currently using mass spectrometry techniques to characterize the interactomes of these isoforms in different organisms.
Characterizing PTM-driven chaperone interactions
Hsp70 interacts with a vast number of proteins in the cell. We are currently pushing the boundaries of mass spectrometry to understand how PTMs on both chaperones and their clients alter their interactions on a global scale.
Using CRISPR-Cas9 to study chaperone function
In the Truman Lab we are currently using CRISPR to:
manipulate chaperone/co-chaperone function
epitope tag chaperones for proteomic studies
randomly mutate chaperones in combination with anticancer drug screens.
If you are interested in collaborating with us on a CRISPR project, please email Dr. Andy Truman at: atruman1 at uncc.edu, we would be happy to help!