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Originally published In Press as doi:10.1074/jbc.M412304200 on December 28, 2004

J. Biol. Chem., Vol. 280, Issue 10, 9439-9449, March 11, 2005
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Interaction of Microtubule-associated Protein-2 and p63

A NEW LINK BETWEEN MICROTUBULES AND ROUGH ENDOPLASMIC RETICULUM MEMBRANES IN NEURONS*

Carole Abi Farah{ddagger}§, Dalinda Liazoghli{ddagger}§, Sébastien Perreault{ddagger}, Mylène Desjardins{ddagger}||, Alain Guimont{ddagger}, Angela Anton{ddagger}, Michel Lauzon{ddagger}, Gert Kreibich**, Jacques Paiement{ddagger}, and Nicole Leclerc, Scholar of Fonds de la Recherche en Santé du Québec{ddagger}{ddagger}{ddagger}

From the {ddagger}Département de Pathologie et Biologie Cellulaire, Université de Montréal, C.P. 6128, Succ. Centre-ville, Montréal, Québec H3C 3J7, Canada and the **Department of Cell Biology, New York University School of Medicine, New York, New York 10016

Neurons are polarized cells presenting two distinct compartments, dendrites and an axon. Dendrites can be distinguished from the axon by the presence of rough endoplasmic reticulum (RER). The mechanism by which the structure and distribution of the RER is maintained in these cells is poorly understood. In the present study, we investigated the role of the dendritic microtubule-associated protein-2 (MAP2) in the RER membrane positioning by comparing their distribution in brain subcellular fractions and in primary hippocampal cells and by examining the MAP2-microtubule interaction with RER membranes in vitro. Subcellular fractionation of rat brain revealed a high MAP2 content in a subfraction enriched with the endoplasmic reticulum markers ribophorin and p63. Electron microscope morphometry confirmed the enrichment of this subfraction with RER membranes. In cultured hippocampal neurons, MAP2 and p63 were found to concomitantly compartmentalize to the dendritic processes during neuronal differentiation. Protein blot overlays using purified MAP2c protein revealed its interaction with p63, and immunoprecipitation experiments performed in HeLa cells showed that this interaction involves the projection domain of MAP2. In an in vitro reconstitution assay, MAP2-containing microtubules were observed to bind to RER membranes in contrast to microtubules containing tau, the axonal MAP. This binding of MAP2c microtubules was reduced when an anti-p63 antibody was added to the assay. The present results suggest that MAP2 is involved in the association of RER membranes with microtubules and thereby could participate in the differential distribution of RER membranes within a neuron.


Received for publication, October 29, 2004 , and in revised form, December 15, 2004.

* This work was supported by a National Sciences and Engineering Research Council of Canada grant and by the Canadian Institute of Health Research Grants MOP-53218 (to N. L.) and MOP-44022 (to J. P.). The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

§ Both authors contributed equally to this work.

Recipient of a studentship from the Centre de Recherche en Sciences Neurologiques.

|| Recipient of a studentship from the National Sciences and Engineering Research Council of Canada.

{ddagger}{ddagger} To whom correspondence should be addressed. Tel.: 514-343-5657; Fax: 514-343-5755; E-mail: nicole.leclerc{at}umontreal.ca.


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