PMID:14752510

Citation	Gotoh, T, Terada, K, Oyadomari, S and Mori, M (2004) hsp70-DnaJ chaperone pair prevents nitric oxide- and CHOP-induced apoptosis by inhibiting translocation of Bax to mitochondria. Cell Death Differ. 11:390-402
Abstract	We reported that the endoplasmic reticulum (ER) stress pathway involving CHOP, a member of the C/EBP transcription factor family, plays a key role in nitric oxide (NO)-mediated apoptosis of macrophages and pancreatic beta cells. We also showed that the cytosolic chaperone pair of hsp70 and dj1 (hsp40/hdj-1) or dj2 (HSDJ/hdj-2) prevents NO-mediated apoptosis upstream of cytochrome c release from mitochondria. To analyze roles of the chaperone pair in preventing apoptosis, RAW 264.7 macrophages stably expressing hsp70 and dj1 or dj2 were established. The chaperone pair prevented LPS/IFN-gamma-induced and NO-mediated apoptosis downstream of CHOP induction. hsp70 mutant protein lacking the ATPase domain or the C-terminal EEVD sequence were not effective in preventing CHOP-induced apoptosis. A mutant dj2 lacking the C-terminal prenylation CaaX motif, was also not effective. When wild-type RAW 264.7 cells were treated with LPS/IFN-gamma, NO-mediated apoptosis was induced, and proapoptotic Bcl-2 family protein Bax was translocated from cytosol to mitochondria. This translocation was prevented in cells stably expressing hsp70/dj2, and in CHOP knockout cells. Overexpression of CHOP in wild-type cells also induced translocation of Bax and this translocation was prevented in cells expressing hsp70/dj2. CHOP-induced apoptosis was prevented by Bax knock-down. Coimmunoprecipitation experiments showed that Bax interacts with both hsp70 and dj1/dj2. ATPase domain of hsp70 was necessary for the binding with Bax. These findings indicate that CHOP-induced apoptosis is mediated by translocation of Bax from the cytosol to the mitochondria, and hsp70/dj1 or dj2 chaperone pair prevents apoptosis by interacting with Bax and preventing translocation to the mitochondria.
Links	PubMed Online version:10.1038/sj.cdd.4401369
Keywords	Adenosine Triphosphatases/metabolism; Animals; Apoptosis/physiology; CCAAT-Enhancer-Binding Proteins/antagonists & inhibitors; CCAAT-Enhancer-Binding Proteins/metabolism; COS Cells; Cell Line; Cercopithecus aethiops; Cytosol/metabolism; HSP40 Heat-Shock Proteins; HSP70 Heat-Shock Proteins/genetics; HSP70 Heat-Shock Proteins/physiology; Heat-Shock Proteins/genetics; Heat-Shock Proteins/physiology; Macrophages/cytology; Macrophages/metabolism; Mice; Mice, Knockout; Mitochondria/metabolism; Molecular Chaperones/metabolism; Molecular Chaperones/pharmacology; Nitric Oxide/antagonists & inhibitors; Protein Transport/physiology; Proto-Oncogene Proteins/antagonists & inhibitors; Proto-Oncogene Proteins/metabolism; Proto-Oncogene Proteins c-bcl-2; Transcription Factor CHOP; Transcription Factors/antagonists & inhibitors; Transcription Factors/metabolism; Transfection; bcl-2-Associated X Protein
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Annotations

Gene product	Qualifier	GO Term	Evidence Code	with/from	Aspect	Extension	Notes	Status
HUMAN:ATF6A	involved_in	GO:0043065: positive regulation of apoptotic process	ECO:0000314: direct assay evidence used in manual assertion		P		Seeded From UniProt	complete
HUMAN:DNJA1	enables	GO:0030957: Tat protein binding	ECO:0000353: physical interaction evidence used in manual assertion	UniProtKB:Q07813	F	has_input:(UniProtKB:Q07812)	Seeded From UniProt	complete
HUMAN:DNJA1	involved_in	GO:1905259: negative regulation of nitrosative stress-induced intrinsic apoptotic signaling pathway	ECO:0000314: direct assay evidence used in manual assertion		P	occurs_in:(CL:0000235)	Seeded From UniProt	complete
HUMAN:DNJA1	involved_in	GO:1903748: negative regulation of establishment of protein localization to mitochondrion	ECO:0000314: direct assay evidence used in manual assertion		P	part_of:(GO:1905259) has_input:(UniProtKB:Q07812)	Seeded From UniProt	complete
MOUSE:DDIT3	involved_in	GO:1990442: intrinsic apoptotic signaling pathway in response to nitrosative stress	ECO:0000314: direct assay evidence used in manual assertion		P		Seeded From UniProt	complete
MOUSE:DDIT3	involved_in	GO:0072655: establishment of protein localization to mitochondrion	ECO:0000314: direct assay evidence used in manual assertion		P	transports_or_maintains_localization_of:(UniProtKB:Q07813) part_of:(GO:0043065)	Seeded From UniProt	complete
MOUSE:DDIT3	part_of	GO:0005634: nucleus	ECO:0000314: direct assay evidence used in manual assertion		C	part_of:(CL:0000235)	Seeded From UniProt	complete
MOUSE:DNJA1	enables	GO:0005515: protein binding	ECO:0000353: physical interaction evidence used in manual assertion	UniProtKB:Q07813	F	happens_during:(GO:0033554)	Seeded From UniProt	complete
MOUSE:BAX	enables	GO:0030544: Hsp70 protein binding	ECO:0000314: direct assay evidence used in manual assertion		F	happens_during:(GO:0033554)	Seeded From UniProt	complete
MOUSE:BAX	enables	GO:0051087: chaperone binding	ECO:0000353: physical interaction evidence used in manual assertion	UniProtKB:P63037	F	happens_during:(GO:0033554)	Seeded From UniProt	complete
MOUSE:BAX	enables	GO:0051087: chaperone binding	ECO:0000353: physical interaction evidence used in manual assertion	UniProtKB:Q99LX0	F	happens_during:(GO:0033554)	Seeded From UniProt	complete
MOUSE:BAX	enables	GO:0005515: protein binding	ECO:0000353: physical interaction evidence used in manual assertion	UniProtKB:P31689	F	has_input:(UniProtKB:P63037)	Seeded From UniProt	complete
MOUSE:BAX	part_of	GO:0005739: mitochondrion	ECO:0000314: direct assay evidence used in manual assertion		C	exists_during:(GO:0006915)	Seeded From UniProt	complete
HUMAN:PARK7	involved_in	GO:1905259: negative regulation of nitrosative stress-induced intrinsic apoptotic signaling pathway	ECO:0000314: direct assay evidence used in manual assertion		P	occurs_in:(CL:0000235)	Seeded From UniProt	complete
MOUSE:PARK7	enables	GO:0005515: protein binding	ECO:0000353: physical interaction evidence used in manual assertion	UniProtKB:Q07813	F	happens_during:(GO:0033554)	Seeded From UniProt	complete
HUMAN:PARK7	involved_in	GO:1905259: negative regulation of nitrosative stress-induced intrinsic apoptotic signaling pathway	ECO:0000314: direct assay evidence used in manual assertion		P	occurs_in:(CL:0000235)	Seeded From UniProt	complete
MOUSE:BAX	part_of	GO:0005737: cytoplasm	ECO:0000314: direct assay evidence used in manual assertion		C		Seeded From UniProt	complete
MOUSE:BAX	part_of	GO:0005739: mitochondrion	ECO:0000314: direct assay evidence used in manual assertion		C	exists_during:(GO:0006915)	Seeded From UniProt	complete
MOUSE:BAX	enables	GO:0030544: Hsp70 protein binding	ECO:0000314: direct assay evidence used in manual assertion		F	happens_during:(GO:0033554)	Seeded From UniProt	complete
MOUSE:BAX	enables	GO:0051087: chaperone binding	ECO:0000353: physical interaction evidence used in manual assertion	UniProtKB:Q99LX0	F	happens_during:(GO:0033554)	Seeded From UniProt	complete
MOUSE:BAX	enables	GO:0051087: chaperone binding	ECO:0000353: physical interaction evidence used in manual assertion	UniProtKB:P63037	F	happens_during:(GO:0033554)	Seeded From UniProt	complete
HUMAN:DNJA1	enables	GO:0051087: chaperone binding	ECO:0000314: direct assay evidence used in manual assertion		F		Seeded From UniProt	complete
HUMAN:DNJA1	involved_in	GO:0070585: protein localization to mitochondrion	ECO:0000315: mutant phenotype evidence used in manual assertion		P		Seeded From UniProt	complete
HUMAN:DNJA1	involved_in	GO:0043066: negative regulation of apoptotic process	ECO:0000315: mutant phenotype evidence used in manual assertion		P		Seeded From UniProt	complete
HUMAN:DNJA1	enables	GO:0030544: Hsp70 protein binding	ECO:0000314: direct assay evidence used in manual assertion		F		Seeded From UniProt	complete
HUMAN:DNJA1	involved_in	GO:0051223: regulation of protein transport	ECO:0000314: direct assay evidence used in manual assertion		P		Seeded From UniProt	complete
HUMAN:DNJA1	involved_in	GO:1905259: negative regulation of nitrosative stress-induced intrinsic apoptotic signaling pathway	ECO:0000314: direct assay evidence used in manual assertion		P	occurs_in:(CL:0000235)	Seeded From UniProt	complete
HUMAN:DNJA1	involved_in	GO:1903748: negative regulation of establishment of protein localization to mitochondrion	ECO:0000314: direct assay evidence used in manual assertion		P	part_of:(GO:1905259) has_regulation_target:(UniProtKB:Q07812)	Seeded From UniProt	complete
HUMAN:DNJA1	enables	GO:0030957: Tat protein binding	ECO:0000353: physical interaction evidence used in manual assertion	UniProtKB:Q07813	F	has_input:(UniProtKB:Q07812)	Seeded From UniProt	complete

References

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