Evaluation of the role of the Endoplasmic Reticulum-Golgi transit in the biogenesis of peroxisomal membrane proteins in wild type and peroxisome biogenesis mutant CHO cells

Peroxisomes are thought to be formed by division of pre-existing peroxisomes after the import of newly synthesized proteins. However, it has been recently suggested that the endoplasmic reticulum (ER) provides an alternative de novo mechanism for peroxisome biogenesis in some cells. To test a possib...

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Authors: TORO,ANDRÉS, ARREDONDO,CRISTIAN, CORDOVA,GONZALO, ARAYA,CLAUDIA, PALACIOS,JOSÉ L, VENEGAS,ALEJANDRO, MORITA,MASASHI, IMANAKA,TSUNEO, SANTOS,MANUEL J
Format: article
Status:Published version
Publication Date:2007
Country:Chile
Institution:CONICYT Chile
Repository:SciELO Chile
OAI Identifier:oai:scielo:S0716-97602007000200014
Online Access:http://www.scielo.cl/scielo.php?script=sci_arttext&pid=S0716-97602007000200014
Access Level:Open access
Keyword:adrenoleukodystrophy
ALDRP
endoplasmic reticulum
peroxisome biogenesis
Pex3p and PMP70
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spelling Evaluation of the role of the Endoplasmic Reticulum-Golgi transit in the biogenesis of peroxisomal membrane proteins in wild type and peroxisome biogenesis mutant CHO cells TORO,ANDRÉS ARREDONDO,CRISTIAN CORDOVA,GONZALO ARAYA,CLAUDIA PALACIOS,JOSÉ L VENEGAS,ALEJANDRO MORITA,MASASHI IMANAKA,TSUNEO SANTOS,MANUEL J adrenoleukodystrophy ALDRP endoplasmic reticulum peroxisome biogenesis Pex3p and PMP70 Peroxisomes are thought to be formed by division of pre-existing peroxisomes after the import of newly synthesized proteins. However, it has been recently suggested that the endoplasmic reticulum (ER) provides an alternative de novo mechanism for peroxisome biogenesis in some cells. To test a possible role of the ER-Golgi transit in peroxisome biogenesis in mammalian cells, we evaluated the biogenesis of three peroxisomal membrane proteins (PMPs): ALDRP (adrenoleukodystrophy related protein), PMP70 and Pex3p in CHO cells. We constructed chimeric genes encoding these PMPs and green fluorescent protein (GFP), and transiently transfected them to wild type and mutant CHO cells, in which normal peroxisomes were replaced by peroxisomal membrane ghosts. The expressed proteins were targeted to peroxisomes and peroxisomal ghosts correctly in the presence or absence of Brefeldin A (BFA), a drug known to block the ER-Golgi transit. Furthermore, low temperature did not disturb the targeting of Pex3p-GFP to peroxisomes. We also constructed two chimeric proteins of PMPs containing an ER retention signal "DEKKMP": GFP-ALDRP-DEKKMP and myc- Pex3p-DEKKMP. These proteins were mostly targeted to peroxisomes. No colocalization with an ER maker was found. These results suggest that the classical ER-Golgi pathway does not play a major role in the biogenesis of mammalian PMPs Sociedad de Biología de Chile http://www.scielo.cl/scielo.php?script=sci_arttext&pid=S0716-97602007000200014
title Evaluation of the role of the Endoplasmic Reticulum-Golgi transit in the biogenesis of peroxisomal membrane proteins in wild type and peroxisome biogenesis mutant CHO cells
spellingShingle Evaluation of the role of the Endoplasmic Reticulum-Golgi transit in the biogenesis of peroxisomal membrane proteins in wild type and peroxisome biogenesis mutant CHO cells
TORO,ANDRÉS
adrenoleukodystrophy
ALDRP
endoplasmic reticulum
peroxisome biogenesis
Pex3p and PMP70
title_short Evaluation of the role of the Endoplasmic Reticulum-Golgi transit in the biogenesis of peroxisomal membrane proteins in wild type and peroxisome biogenesis mutant CHO cells
title_full Evaluation of the role of the Endoplasmic Reticulum-Golgi transit in the biogenesis of peroxisomal membrane proteins in wild type and peroxisome biogenesis mutant CHO cells
title_fullStr Evaluation of the role of the Endoplasmic Reticulum-Golgi transit in the biogenesis of peroxisomal membrane proteins in wild type and peroxisome biogenesis mutant CHO cells
title_full_unstemmed Evaluation of the role of the Endoplasmic Reticulum-Golgi transit in the biogenesis of peroxisomal membrane proteins in wild type and peroxisome biogenesis mutant CHO cells
title_sort Evaluation of the role of the Endoplasmic Reticulum-Golgi transit in the biogenesis of peroxisomal membrane proteins in wild type and peroxisome biogenesis mutant CHO cells
author TORO,ANDRÉS
author_facet TORO,ANDRÉS
ARREDONDO,CRISTIAN
CORDOVA,GONZALO
ARAYA,CLAUDIA
PALACIOS,JOSÉ L
VENEGAS,ALEJANDRO
MORITA,MASASHI
IMANAKA,TSUNEO
SANTOS,MANUEL J
author_role author
author2 ARREDONDO,CRISTIAN
CORDOVA,GONZALO
ARAYA,CLAUDIA
PALACIOS,JOSÉ L
VENEGAS,ALEJANDRO
MORITA,MASASHI
IMANAKA,TSUNEO
SANTOS,MANUEL J
author2_role author
author
author
author
author
author
author
author
topic adrenoleukodystrophy
ALDRP
endoplasmic reticulum
peroxisome biogenesis
Pex3p and PMP70
topic_facet adrenoleukodystrophy
ALDRP
endoplasmic reticulum
peroxisome biogenesis
Pex3p and PMP70
description Peroxisomes are thought to be formed by division of pre-existing peroxisomes after the import of newly synthesized proteins. However, it has been recently suggested that the endoplasmic reticulum (ER) provides an alternative de novo mechanism for peroxisome biogenesis in some cells. To test a possible role of the ER-Golgi transit in peroxisome biogenesis in mammalian cells, we evaluated the biogenesis of three peroxisomal membrane proteins (PMPs): ALDRP (adrenoleukodystrophy related protein), PMP70 and Pex3p in CHO cells. We constructed chimeric genes encoding these PMPs and green fluorescent protein (GFP), and transiently transfected them to wild type and mutant CHO cells, in which normal peroxisomes were replaced by peroxisomal membrane ghosts. The expressed proteins were targeted to peroxisomes and peroxisomal ghosts correctly in the presence or absence of Brefeldin A (BFA), a drug known to block the ER-Golgi transit. Furthermore, low temperature did not disturb the targeting of Pex3p-GFP to peroxisomes. We also constructed two chimeric proteins of PMPs containing an ER retention signal "DEKKMP": GFP-ALDRP-DEKKMP and myc- Pex3p-DEKKMP. These proteins were mostly targeted to peroxisomes. No colocalization with an ER maker was found. These results suggest that the classical ER-Golgi pathway does not play a major role in the biogenesis of mammalian PMPs
publishDate 2007
format article
status_str publishedVersion
url http://www.scielo.cl/scielo.php?script=sci_arttext&pid=S0716-97602007000200014
eu_rights_str_mv openAccess
publisher Sociedad de Biología de Chile
institution CONICYT
collection SciELO Chile
reponame_str SciELO Chile
instname_str CONICYT Chile
_version_ 1878404558038761472
publishDateSort 2007
author_browse ARAYA,CLAUDIA
ARREDONDO,CRISTIAN
CORDOVA,GONZALO
IMANAKA,TSUNEO
MORITA,MASASHI
PALACIOS,JOSÉ L
SANTOS,MANUEL J
TORO,ANDRÉS
VENEGAS,ALEJANDRO
publisherStr Sociedad de Biología de Chile
score 6,924472