Nucleocytoplasmic trafficking of the molecular chaperone hsp104 in unstressed and heat-shocked cells

作者：Tkach Johnny M; Glover John R^*

来源：Traffic, 2008, 9(1): 39-56.

DOI：10.1111/j.1600-0854.2007.00666.x

• budding yeast
• heat-shock protein
• molecular chaperone
• protein folding
• protein trafficking
• thermotolerance

Hsp104 is a molecular chaperone in yeast that restores solubility and activity to inactivated proteins after severe heat shock. We investigated the mechanisms that influence Hsp104 subcellular distribution in both unstressed and heat-shocked cells. In unstressed cells, Hsp104 and a green fluorescent protein-Hsp104 fusion protein were detected in both the nucleus and the cytoplasm. We demonstrate that a 17-amino-acid sequence of Hsp104 nuclear localization sequence 17 (NLS17) is sufficient to target a reporter molecule to the nucleus and is also necessary for normal Hsp104 subcellular distribution. The nuclear targeting function of NLS17 is genetically dependent on KAP95 and KAP121. In addition, wild-type Hsp104, but not an NLS17-mutated Hsp104 variant, accumulated in the nucleus of cells depleted for the general export factor Xpo1. Interestingly, severe, nonlethal heat shock enhances the nuclear levels of Hsp104 in an NLS17-independent manner. Under these conditions, we demonstrate that karyopherin-mediated nuclear transport is impaired, while the integrity of the nuclear-cytoplasmic barrier remains intact. Based on these observations, we propose that Hsp104 continues to access the nucleus during severe heat shock using a karyopherin-independent mechanism.

• 出版日期2008-1