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

Yoshinori Kashiwayama, Midori Seki, Akina Yasui, Yoshiyuki Murasaki, Masashi Morita, Yukari Yamashita, Masao Sakaguchi, Yoshitaka Tanaka, Tsuneo Imanaka

Research output: Contribution to journalArticle

33 Citations (Scopus)

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 NH2-terminal deletion constructs of P70R. Among the NH2-terminal deletion constructs, mutant proteins starting with hydrophobic transmembrane segment (TMS) were localized to ER, but the ones containing the NH2-terminal hydrophilic cytosolic domain were not. ABC subfamily D proteins destined for peroxisomes have NH2-terminal hydrophilic region adjacent to TMS1. However, only P70R lacks the region and is translated with NH2-terminal hydrophobic TMS1. Furthermore, attachment of the NH2-terminal hydrophilic domain to the NH2-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 NH2-terminal region determines the subcellular localization of ABC subfamily D proteins.

Original languageEnglish
Pages (from-to)190-205
Number of pages16
JournalExperimental Cell Research
Volume315
Issue number2
DOIs
Publication statusPublished - Jan 15 2009

Fingerprint

Peroxisomes
Endoplasmic Reticulum
Membrane Proteins
Adenosine Triphosphate
Proteins
Carrier Proteins
CHO Cells
Mutant Proteins
Fluorescent Antibody Technique
Membranes

All Science Journal Classification (ASJC) codes

  • Cell Biology

Cite this

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. / Kashiwayama, Yoshinori; Seki, Midori; Yasui, Akina; Murasaki, Yoshiyuki; Morita, Masashi; Yamashita, Yukari; Sakaguchi, Masao; Tanaka, Yoshitaka; Imanaka, Tsuneo.

In: Experimental Cell Research, Vol. 315, No. 2, 15.01.2009, p. 190-205.

Research output: Contribution to journalArticle

Kashiwayama, Yoshinori ; Seki, Midori ; Yasui, Akina ; Murasaki, Yoshiyuki ; Morita, Masashi ; Yamashita, Yukari ; Sakaguchi, Masao ; Tanaka, Yoshitaka ; Imanaka, Tsuneo. / 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. In: Experimental Cell Research. 2009 ; Vol. 315, No. 2. pp. 190-205.
@article{c6acfde9073642cea6d79c5a68fcbc3b,
title = "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",
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 NH2-terminal deletion constructs of P70R. Among the NH2-terminal deletion constructs, mutant proteins starting with hydrophobic transmembrane segment (TMS) were localized to ER, but the ones containing the NH2-terminal hydrophilic cytosolic domain were not. ABC subfamily D proteins destined for peroxisomes have NH2-terminal hydrophilic region adjacent to TMS1. However, only P70R lacks the region and is translated with NH2-terminal hydrophobic TMS1. Furthermore, attachment of the NH2-terminal hydrophilic domain to the NH2-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 NH2-terminal region determines the subcellular localization of ABC subfamily D proteins.",
author = "Yoshinori Kashiwayama and Midori Seki and Akina Yasui and Yoshiyuki Murasaki and Masashi Morita and Yukari Yamashita and Masao Sakaguchi and Yoshitaka Tanaka and Tsuneo Imanaka",
year = "2009",
month = "1",
day = "15",
doi = "10.1016/j.yexcr.2008.10.031",
language = "English",
volume = "315",
pages = "190--205",
journal = "Experimental Cell Research",
issn = "0014-4827",
publisher = "Academic Press Inc.",
number = "2",

}

TY - JOUR

T1 - 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

AU - Kashiwayama, Yoshinori

AU - Seki, Midori

AU - Yasui, Akina

AU - Murasaki, Yoshiyuki

AU - Morita, Masashi

AU - Yamashita, Yukari

AU - Sakaguchi, Masao

AU - Tanaka, Yoshitaka

AU - Imanaka, Tsuneo

PY - 2009/1/15

Y1 - 2009/1/15

N2 - 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 NH2-terminal deletion constructs of P70R. Among the NH2-terminal deletion constructs, mutant proteins starting with hydrophobic transmembrane segment (TMS) were localized to ER, but the ones containing the NH2-terminal hydrophilic cytosolic domain were not. ABC subfamily D proteins destined for peroxisomes have NH2-terminal hydrophilic region adjacent to TMS1. However, only P70R lacks the region and is translated with NH2-terminal hydrophobic TMS1. Furthermore, attachment of the NH2-terminal hydrophilic domain to the NH2-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 NH2-terminal region determines the subcellular localization of ABC subfamily D proteins.

AB - 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 NH2-terminal deletion constructs of P70R. Among the NH2-terminal deletion constructs, mutant proteins starting with hydrophobic transmembrane segment (TMS) were localized to ER, but the ones containing the NH2-terminal hydrophilic cytosolic domain were not. ABC subfamily D proteins destined for peroxisomes have NH2-terminal hydrophilic region adjacent to TMS1. However, only P70R lacks the region and is translated with NH2-terminal hydrophobic TMS1. Furthermore, attachment of the NH2-terminal hydrophilic domain to the NH2-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 NH2-terminal region determines the subcellular localization of ABC subfamily D proteins.

UR - http://www.scopus.com/inward/record.url?scp=57749175975&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=57749175975&partnerID=8YFLogxK

U2 - 10.1016/j.yexcr.2008.10.031

DO - 10.1016/j.yexcr.2008.10.031

M3 - Article

C2 - 19010322

AN - SCOPUS:57749175975

VL - 315

SP - 190

EP - 205

JO - Experimental Cell Research

JF - Experimental Cell Research

SN - 0014-4827

IS - 2

ER -