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PMID:19010322

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Citation

Kashiwayama, Y, Seki, M, Yasui, A, Murasaki, Y, Morita, M, Yamashita, Y, Sakaguchi, M, Tanaka, Y and Imanaka, T (2009) 70-kDa peroxisomal membrane protein related protein (P70R/ABCD4) localizes to endoplasmic reticulum not peroxisomes, and NH2-terminal hydrophobic property determines the subcellular localization of ABC subfamily D proteins. Exp. Cell Res. 315:190-205

Abstract

70-kDa peroxisomal membrane protein related protein (P70R/ABCD4) is a member of ATP-binding cassette (ABC) protein subfamily D. ABC subfamily D proteins are also known as peroxisomal ABC proteins. Therefore, P70R is thought to be a peroxisomal membrane protein. However, the subcellular localization of P70R is not extensively investigated. In this study, we transiently expressed P70R in fusion with HA (P70R-HA) in CHO cells and examined subcellular localization by immunofluorescence. Surprisingly, P70R-HA was localized to the endoplasmic reticulum (ER), not to peroxisomes. To examine the ER-targeting property of P70R, we expressed various NH(2)-terminal deletion constructs of P70R. Among the NH(2)-terminal deletion constructs, mutant proteins starting with hydrophobic transmembrane segment (TMS) were localized to ER, but the ones containing the NH(2)-terminal hydrophilic cytosolic domain were not. ABC subfamily D proteins destined for peroxisomes have NH(2)-terminal hydrophilic region adjacent to TMS1. However, only P70R lacks the region and is translated with NH(2)-terminal hydrophobic TMS1. Furthermore, attachment of the NH(2)-terminal hydrophilic domain to the NH(2)-terminus of P70R excluded P70R from the ER-targeting pathway. These data suggest that P70R resides in the ER but not the peroxisomal membranes, and the hydrophobic property of NH(2)-terminal region determines the subcellular localization of ABC subfamily D proteins.

Links

PubMed Online version:10.1016/j.yexcr.2008.10.031

Keywords

ATP-Binding Cassette Transporters/genetics; ATP-Binding Cassette Transporters/metabolism; Animals; CHO Cells; COS Cells; Cercopithecus aethiops; Cricetinae; Cricetulus; Endoplasmic Reticulum/metabolism; Glycosylation; Intracellular Membranes/metabolism; Liver/metabolism; Male; Mannosyl-Glycoprotein Endo-beta-N-Acetylglucosaminidase/metabolism; Peroxisomes/metabolism; Protein Binding; Protein Sorting Signals/physiology; Protein Transport/physiology; Rats; Rats, Wistar; Sequence Deletion; Transfection

Significance

Annotations

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

HUMAN:ABCD4

NOT|located_in

GO:0005777: peroxisome

ECO:0000314: direct assay evidence used in manual assertion

C

Seeded From UniProt

complete

MOUSE:ABCD4

located_in

GO:0005789: endoplasmic reticulum membrane

ECO:0000314: direct assay evidence used in manual assertion

C

Seeded From UniProt

complete

MOUSE:ABCD4

NOT|located_in

GO:0005777: peroxisome

ECO:0000314: direct assay evidence used in manual assertion

C

Seeded From UniProt

complete

RAT:ABCD3

located_in

GO:0005739: mitochondrion

ECO:0000314: direct assay evidence used in manual assertion

C

Seeded From UniProt

complete

MOUSE:ABCD3

enables

GO:0005515: protein binding

ECO:0000353: physical interaction evidence used in manual assertion

UniProtKB:Q8VCI5

F

Seeded From UniProt

complete

MOUSE:ABCD3

located_in

GO:0005777: peroxisome

ECO:0000314: direct assay evidence used in manual assertion

C

Seeded From UniProt

complete

RAT:Q5FWT4

located_in

GO:0005789: endoplasmic reticulum membrane

ECO:0000314: direct assay evidence used in manual assertion

C

Seeded From UniProt

complete

MOUSE:PEX19

enables

GO:0005515: protein binding

ECO:0000353: physical interaction evidence used in manual assertion

UniProtKB:P55096

F

Seeded From UniProt

complete

RAT:Q5FWT4

part_of

GO:0005789: endoplasmic reticulum membrane

ECO:0000314: direct assay evidence used in manual assertion

C

Seeded From UniProt

complete


See also

References

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