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PMID:15623521

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Citation

Farah, CA, Liazoghli, D, Perreault, S, Desjardins, M, Guimont, A, Anton, A, Lauzon, M, Kreibich, G, Paiement, J and Leclerc, N (2005) Interaction of microtubule-associated protein-2 and p63: a new link between microtubules and rough endoplasmic reticulum membranes in neurons. J. Biol. Chem. 280:9439-49

Abstract

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.

Links

PubMed Online version:10.1074/jbc.M412304200

Keywords

Animals; Brain/embryology; Brain/metabolism; Cattle; Cell Line; Embryo, Mammalian; Embryo, Nonmammalian; Endoplasmic Reticulum/physiology; Endoplasmic Reticulum/ultrastructure; Golgi Apparatus/metabolism; Golgi Apparatus/ultrastructure; Hippocampus/physiology; Intracellular Membranes/metabolism; Microscopy, Electron; Microsomes/metabolism; Microsomes/ultrastructure; Microtubule-Associated Proteins/metabolism; Microtubules/metabolism; Microtubules/ultrastructure; Neurons/physiology; Rats; Rats, Sprague-Dawley; Recombinant Proteins/metabolism; Spodoptera; Transcription Factors/metabolism; Transfection

Significance

Annotations

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

RAT:RPN1

located_in

GO:0005791: rough endoplasmic reticulum

ECO:0000314: direct assay evidence used in manual assertion

C

Seeded From UniProt

complete

RAT:MTAP2

enables

GO:0005515: protein binding

ECO:0000353: physical interaction evidence used in manual assertion

RGD:620863

F

Seeded From UniProt

complete

RAT:MTAP2

located_in

GO:0005791: rough endoplasmic reticulum

ECO:0000314: direct assay evidence used in manual assertion

C

Seeded From UniProt

complete

RAT:MTAP2

located_in

GO:0043005: neuron projection

ECO:0000314: direct assay evidence used in manual assertion

C

Seeded From UniProt

complete

RAT:MTAP2

part_of

GO:0032991: protein-containing complex

ECO:0000314: direct assay evidence used in manual assertion

C

Seeded From UniProt

complete

RAT:MTAP2

located_in

GO:0030425: dendrite

ECO:0000314: direct assay evidence used in manual assertion

C

Seeded From UniProt

complete

RAT:MTAP2

located_in

GO:0005790: smooth endoplasmic reticulum

ECO:0000314: direct assay evidence used in manual assertion

C

Seeded From UniProt

complete

RAT:TAU

located_in

GO:0043005: neuron projection

ECO:0000314: direct assay evidence used in manual assertion

C

Seeded From UniProt

complete

RAT:TAU

located_in

GO:0030424: axon

ECO:0000314: direct assay evidence used in manual assertion

C

Seeded From UniProt

complete

RAT:RPN2

located_in

GO:0005791: rough endoplasmic reticulum

ECO:0000314: direct assay evidence used in manual assertion

C

Seeded From UniProt

complete

RAT:P63

enables

GO:0019904: protein domain specific binding

ECO:0000353: physical interaction evidence used in manual assertion

RGD:3044

F

Seeded From UniProt

complete

RAT:P63

located_in

GO:0005634: nucleus

ECO:0000314: direct assay evidence used in manual assertion

C

Seeded From UniProt

complete

RAT:P63

located_in

GO:0005791: rough endoplasmic reticulum

ECO:0000314: direct assay evidence used in manual assertion

C

Seeded From UniProt

complete

RAT:P63

located_in

GO:0043005: neuron projection

ECO:0000314: direct assay evidence used in manual assertion

C

Seeded From UniProt

complete

RAT:P63

part_of

GO:0032991: protein-containing complex

ECO:0000314: direct assay evidence used in manual assertion

C

Seeded From UniProt

complete

RAT:P63

located_in

GO:0030425: dendrite

ECO:0000314: direct assay evidence used in manual assertion

C

Seeded From UniProt

complete

RAT:VDAC3

located_in

GO:0005791: rough endoplasmic reticulum

ECO:0000314: direct assay evidence used in manual assertion

C

Seeded From UniProt

complete

RAT:TAU

part_of

GO:0043005: neuron projection

ECO:0000314: direct assay evidence used in manual assertion

C

Seeded From UniProt

complete

RAT:TAU

part_of

GO:0030424: axon

ECO:0000314: direct assay evidence used in manual assertion

C

Seeded From UniProt

complete


See also

References

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