Multi-chaperone complexes regulate the folding of interferon-gamma in the endoplasmic reticulum.

The quality control mechanisms directing the folding of cytokines in the endoplasmic reticulum (ER) are poorly understood. We have investigated ER chaperone usage by the cytokine interferon-gamma (IFN-gamma). ATP-depletion or inhibition of N-glycosylation was found to cause IFN-gamma to accumulate into detergent-insoluble aggregates in the ER. Six chaperones, GRP94, GRP78, ERp72, PDI, CaBP1/P5 and CRT were found to associate with IFN-gamma during its steady state folding. Interaction of the five first chaperones with IFN-gamma was regulated co-ordinately by ATP. These chaperones were recently reported to be part of a multi-chaperone complex involved in the folding of complex, multi-subunit proteins. Our data suggest that also proteins with a relatively simple quaternary structure such as cytokines may fold in association with this complex. In addition, we identified calreticulin as the major chaperone interacting with IFN-gamma, and the related class II cytokine interleukin-10, during heat-shock in vivo. IFN-gamma was maintained in a folding-competent form by calreticulin during heat-shock and released during subsequent recovery at 37 degrees C. This interaction was observed in both recombinant (CHO-F11) and natural producer cells (Jurkat, NK-92MI) of IFN-gamma. Since cytokines such as IFN-gamma and IL-10 are frequently produced in the course of inflammatory conditions associated with fever, the thermo-protective effect of calreticulin may constitute a previously unrecognized component of the cellular cytokine production machinery, of likely relevance in sustaining cytokine folding and secretion in pathophysiological conditions.