Functional organization of microtubule-associated protein tau. Identification of regions which affect microtubule growth, nucleation, and bundle formation in vitro

Roland Brandt; Gloria Lee

doi:10.1016/S0021-9258(18)53710-8

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Functional organization of microtubule-associated protein tau. Identification of regions which affect microtubule growth, nucleation, and bundle formation in vitro

Journal article

Open access

Peer reviewed

Functional organization of microtubule-associated protein tau. Identification of regions which affect microtubule growth, nucleation, and bundle formation in vitro

Roland Brandt and Gloria Lee

The Journal of biological chemistry, Vol.268(5), pp.3414-3419

02/15/1993

DOI: 10.1016/S0021-9258(18)53710-8

PMID: 8429017

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Abstract

Tau protein is a microtubule-associated protein that is almost exclusively expressed in the brain and is enriched in the axon. Determination of tau's sequence has revealed three to four tandem repeats that have been shown to constitute the microtubule binding site. In order to study the functional organization of tau, we prepared a series of truncated tau fragments using an Escherichia coli expression system. We assayed each fragment's activity in promoting growth of microtubules and in nucleating free microtubules. We found that tau's ability to nucleate microtubules requires the presence of additional sequence amino-terminal to that required for growth. We demonstrate that tau's carboxyl and amino termini differentially affect microtubule growth and nucleation. Finally, we show that in vitro microtubule bundle formation occurs when tubulin is assembled in the presence of an amino- and carboxyl-terminally truncated tau protein, whereas almost no bundling is observed in the presence of full-length tau or tau fragments that contain the amino terminus in addition to the repeat domain. We conclude that although the presence of the repeat domain promotes the growth of microtubules, the structural requirements for nucleation activity are more stringent. The differentiation between the growth promoting and nucleation activities on the structural level makes it possible for the two activities to be differentially regulated in vivo.

Polymerase Chain Reaction

Recombinant Proteins - metabolism

Enzyme-Linked Immunosorbent Assay

Electrophoresis, Polyacrylamide Gel

tau Proteins - metabolism

Microscopy, Electron

tau Proteins - isolation & purification

Macromolecular Substances

Brain - metabolism

Recombinant Proteins - isolation & purification

Microtubules - metabolism

Tubulin - metabolism

Animals

tau Proteins - genetics

Cattle

Escherichia coli - genetics

Cloning, Molecular

Fluorescent Antibody Technique

Microtubules - ultrastructure

Tubulin - isolation & purification

Details

Title: Subtitle: Functional organization of microtubule-associated protein tau. Identification of regions which affect microtubule growth, nucleation, and bundle formation in vitro
Creators: Roland Brandt - Center for Neurologic Diseases, Brigham and Women's Hospital, Harvard Medical School, Boston, Massachusetts 02115
Gloria Lee
Resource Type: Journal article
Publication Details: The Journal of biological chemistry, Vol.268(5), pp.3414-3419
DOI: 10.1016/S0021-9258(18)53710-8
PMID: 8429017
NLM abbreviation: J Biol Chem
ISSN: 0021-9258
eISSN: 1083-351X
Publisher: United States
Grant note: GM39300 / NIGMS NIH HHS
Language: English
Date published: 02/15/1993
Academic Unit: Iowa Neuroscience Institute; Immunology; Internal Medicine
Record Identifier: 9984065477702771

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