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AtJ3 (J3)—a member of the Arabidopsis cytosolic HSP40 family—harbors a C-terminal CaaX motif for farnesylation, which is exclusively catalyzed by protein farnesyltransferase (PFT). Previously, prolonged incubation at 37 °C for 4 d was found to be lethal to the heat-intolerant 5 (hit5) mutant lacking PFT and transgenic j3 plants expressing a CaaX-abolishing J3C417S construct, indicating that farnesylated J3 is essential for heat tolerance in plants. Given the role of HSP40s as cochaperones of HSP70s, the thermal sensitivity of five individual cytosolic HSP70 (HSP70-1 to HSP70-5) knockout mutants was tested in this study. Only hsp70-4 was sensitive to the prolonged heat treatment like hit5 and j3. The bimolecular fluorescence complementation (BiFC) assay revealed that HSP70-4 interacted with J3 and J3C417S in vivo at normal (23 °C) and high (37 °C) temperatures. At 23 °C, both HSP70-4–J3 and HSP70-4–J3C417S BiFC signals were uniformly distributed across the cell. However, following treatment at 37 °C, HSP70-4–J3, but not HSP70-4–J3C417S, BiFC signals were detected as discernable foci. These heat-induced HSP70-4–J3 BiFC foci were localized in heat stress granules (HSGs). In addition, hsp70-4 and J3C417S accumulated more insoluble proteins than the wild type. Thus, farnesylated J3 dictates the chaperone function of HSP70-4 in HSGs. Collectively, this study identified the first HSP70/HSP40-type chaperone machinery playing a crucial role in protecting plants against prolonged heat stress, and demonstrated the significance of protein farnesylation in its protective function.
- Heat stress granule (HSG)
- HEAT-INTOLERANT 5 (HIT5)
- J-domain protein
- Protein farnesylation
- Protein farnesyltransferase (PFT)
FingerprintDive into the research topics of 'HSP70-4 and farnesylated AtJ3 constitute a specific HSP70/HSP40-based chaperone machinery essential for prolonged heat stress tolerance in Arabidopsis'. Together they form a unique fingerprint.
- 1 Finished
Forward and Reverse Genetics Approaches to Dissect Protein Farnesylation-Mediated Heat Stress Responses in Plants(2/3)
1/08/20 → 31/07/21