title: Quantitative analysis of the interplay between hsc70 and its co-chaperone HspBP1.
abstract: Background. Chaperones and their co-factors are components of a cellular network; they collaborate to maintain proteostasis under normal and harmful conditions. In particular, hsp70 family members and their co-chaperones are essential to repair damaged proteins. Co-chaperones are present in different subcellular compartments, where they modulate chaperone activities. Methods and Results. Our studies assessed the relationship between hsc70 and its co-factor HspBP1 in human cancer cells. HspBP1 promotes nucleotide exchange on hsc70, but has also chaperone-independent functions. We characterized the interplay between hsc70 and HspBP1 by quantitative confocal microscopy combined with automated image analyses and statistical evaluation. Stress and the recovery from insult changed significantly the subcellular distribution of hsc70, but had little effect on HspBP1. Single-cell measurements and regression analysis revealed that the links between the chaperone and its co-factor relied on (i) the physiological state of the cell and (ii) the subcellular compartment. As such, we identified a linear relationship and strong correlation between hsc70 and HspBP1 distribution in control and heat-shocked cells; this correlation changed in a compartment-specific fashion during the recovery from stress. Furthermore, we uncovered significant stress-induced changes in the colocalization between hsc70 and HspBP1 in the nucleus and cytoplasm. Discussion. Our quantitative approach defined novel properties of the co-chaperone HspBP1 as they relate to its interplay with hsc70. We propose that changes in cell physiology promote chaperone redistribution and thereby stimulate chaperone-independent functions of HspBP1.
authors.affiliation: Department of Physiology, McGill University , Montreal, Quebec , Canada.
authors.forename: Hicham
authors.initials: H
authors.lastname: Mahboubi
authors.affiliation: Department of Physiology, McGill University , Montreal, Quebec , Canada.
authors.forename: Ursula
authors.initials: U
authors.lastname: Stochaj
keywords: Chaperone; Co-chaperone; Heat shock protein; Quantitative microscopy; Regression analysis; Single-cell analysis; Stress response; Subcellular protein distribution