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PMID:21446055

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Citation

Suntio, T, Shiryaev, SA and Makarow, M (2011) ATPase activity of a yeast secretory glycoprotein allows ER exit during inactivation of COPII components Sec24p and Sec13p. Yeast 28:453-65

Abstract

Proteins exit the endoplasmic reticulum (ER) in vesicles pinching off from the membrane at sites covered by the COPII coat, which consists of Sec23/24p and Sec13/31p. We have shown that the glycoprotein Hsp150 exits the ER in the absence of Sec13p or any member of the Sec24p family. The determinant responsible for this resides in the C-terminal domain of Hsp150 (CTD). Here, A- and B-type Walker motifs were identified in the CTD. Authentic Hsp150 from the yeast culture medium, as well as Hsp150 and the CTD fragment produced in Escherichia coli, exhibited ATPase activity nearly three times higher than the published activity of the ER chaperone Kar2p/BiP. Deletion of the Walker motif, and a K335A mutation in it, abolished the ATPase activity. Hsp150 homologues Pir3p and Pir4p, differing in critical amino acids of the Walker motif, also lacked ATPase activity. Unexpectedly, inactivation of the ATPase activity blocked ER exit of Hsp150 in the absence of Sec24p or Sec13p function, whereas secretion in normal cells was not compromised. To our knowledge this is the first documentation of the ATPase activity of a protein serving an intracellular transport function.

Links

PubMed Online version:10.1002/yea.1850

Keywords

Adenosine Triphosphatases/metabolism; Amino Acid Motifs; Amino Acid Sequence; Amino Acid Substitution/genetics; Endoplasmic Reticulum/metabolism; Escherichia coli/genetics; Gene Expression; Glycoproteins/genetics; Glycoproteins/isolation & purification; Glycoproteins/metabolism; Heat-Shock Proteins/genetics; Heat-Shock Proteins/isolation & purification; Heat-Shock Proteins/metabolism; Membrane Proteins/deficiency; Membrane Proteins/genetics; Membrane Proteins/metabolism; Molecular Sequence Data; Nuclear Pore Complex Proteins/deficiency; Nuclear Pore Complex Proteins/genetics; Nuclear Pore Complex Proteins/metabolism; Recombinant Proteins/genetics; Recombinant Proteins/isolation & purification; Recombinant Proteins/metabolism; Saccharomyces cerevisiae/genetics; Saccharomyces cerevisiae/metabolism; Saccharomyces cerevisiae Proteins/genetics; Saccharomyces cerevisiae Proteins/isolation & purification; Saccharomyces cerevisiae Proteins/metabolism; Sequence Deletion

Significance

Annotations

Gene product Qualifier GO Term Evidence Code with/from Aspect Extension Notes Status

YEAST:HS150

enables

GO:0016887: ATP hydrolysis activity

ECO:0000314: direct assay evidence used in manual assertion

F

Seeded From UniProt

complete

YEAST:HS150

enables

GO:0016887: ATP hydrolysis activity

ECO:0000315: mutant phenotype evidence used in manual assertion

F

Seeded From UniProt

complete


See also

References

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