Two distinct domains of protein 4.1 critical for assembly of functional nuclei in vitro

Sharon Wald Krauss; Rebecca Heald; Gloria Lee; Wataru Nunomura; J Aura Gimm; Narla Mohandas; Joel Anne Chasis

doi:10.1074/jbc.M204135200

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Two distinct domains of protein 4.1 critical for assembly of functional nuclei in vitro

Journal article

Open access

Peer reviewed

Two distinct domains of protein 4.1 critical for assembly of functional nuclei in vitro

Sharon Wald Krauss, Rebecca Heald, Gloria Lee, Wataru Nunomura, J Aura Gimm, Narla Mohandas and Joel Anne Chasis

The Journal of biological chemistry, Vol.277(46), pp.44339-44346

11/15/2002

DOI: 10.1074/jbc.M204135200

PMID: 12171917

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Abstract

Protein 4.1R, a multifunctional structural protein, acts as an adaptor in mature red cell membrane skeletons linking spectrin-actin complexes to plasma membrane-associated proteins. In nucleated cells protein 4.1 is not associated exclusively with plasma membrane but is also detected at several important subcellular locations crucial for cell division. To identify 4.1 domains having critical functions in nuclear assembly, 4.1 domain peptides were added to Xenopus egg extract nuclear reconstitution reactions. Morphologically disorganized, replication deficient nuclei assembled when spectrin-actin-binding domain or NuMA-binding C-terminal domain peptides were present. However, control variant spectrin-actin-binding domain peptides incapable of binding actin or mutant C-terminal domain peptides with reduced NuMA binding had no deleterious effects on nuclear reconstitution. To test whether 4.1 is required for proper nuclear assembly, 4.1 isoforms were depleted with spectrin-actin binding or C-terminal domain-specific antibodies. Nuclei assembled in the depleted extracts were deranged. However, nuclear assembly could be rescued by the addition of recombinant 4.1R. Our data establish that protein 4.1 is essential for nuclear assembly and identify two distinct 4.1 domains, initially characterized in cytoskeletal interactions, that have crucial and versatile functions in nuclear assembly.

Cytoskeletal Proteins

Protein Structure, Tertiary

Xenopus

Amino Acid Sequence

Exons

Actins - metabolism

Molecular Sequence Data

Neuropeptides

Sequence Homology, Amino Acid

Microscopy, Confocal

Animals

Cell Nucleus - metabolism

Bromodeoxyuridine - pharmacology

Membrane Proteins - chemistry

Protein Binding

Membrane Proteins - metabolism

Mutation

Fluorescent Antibody Technique, Indirect

Microscopy, Fluorescence

Details

Title: Subtitle: Two distinct domains of protein 4.1 critical for assembly of functional nuclei in vitro
Creators: Sharon Wald Krauss - Department of Subcellular Structure, Life Sciences Division, University of California, Lawrence Berkeley National Laboratory, 94720, USA. sakrauss@lbl.gov
Rebecca Heald
Gloria Lee
Wataru Nunomura
J Aura Gimm
Narla Mohandas
Joel Anne Chasis
Resource Type: Journal article
Publication Details: The Journal of biological chemistry, Vol.277(46), pp.44339-44346
DOI: 10.1074/jbc.M204135200
PMID: 12171917
NLM abbreviation: J Biol Chem
ISSN: 0021-9258
eISSN: 1083-351X
Publisher: United States
Grant note: DK32094 / NIDDK NIH HHS GM57839 / NIGMS NIH HHS DK59079 / NIDDK NIH HHS
Language: English
Date published: 11/15/2002
Academic Unit: Iowa Neuroscience Institute; Immunology; Internal Medicine
Record Identifier: 9984070722102771

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