Red blood cell protein 4.1 (4.1R) is an 80- kD erythrocyte phosphoprotein that stabilizes the spectrin/actin cytoskeleton. In nonerythroid cells, multiple 4.1R isoforms arise from a single gene by alternative splicing and predominantly code for a 135-kD isoform. This isoform contains a 209 amino acid extension at its NH2 terminus (head piece; HP). Immunoreactive epitopes specific for HP have been detected within the cell nucleus, nuclear matrix, centrosomes, and parts of the mitotic apparatus in dividing cells. Using a yeast two-hybrid system, in vitro binding assays, coimmunolocalization, and coimmunoprecipitation studies, we show that a 135-kD 4.1R isoform specifically interacts with the nuclear mitotic apparatus (NuMA) protein. NuMA and 4.1R partially colocalize in the interphase nucleus of MDCK cells and redistribute to the spindle poles early in mitosis. Protein 4.1R associates with NuMA in the interphase nucleus and forms a complex with spindle pole organizing proteins, NuMA, dynein, and dynactin during cell division. Overexpression of a 135-kD isoform of 4.1R alters the normal distribution of NuMA in the interphase nucleus. The minimal sequence sufficient for this interaction has been mapped to the amino acids encoded by exons 20 and 21 of 4.1R and residues 1788–1810 of NuMA. Our results not only suggest that 4.1R could, possibly, play an important role in organizing the nuclear architecture, mitotic spindle, and spindle poles, but also could define a novel role for its 22–24-kD domain.
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5 April 1999
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April 05 1999
A Nonerythroid Isoform of Protein 4.1R Interacts with the Nuclear Mitotic Apparatus (NuMA) Protein
Subhendra N. Mattagajasingh,
Subhendra N. Mattagajasingh
*Department of Medicine, The Johns Hopkins University School of Medicine, Baltimore, Maryland 21205; ‡Department of Biology, Yale University, New Haven, Connecticut 06511; §Boyer Center for Molecular Medicine and Department of Pathology, Yale School of Medicine, New Haven, Connecticut 06510; and ‖Department of Molecular Biology and Genetics, The Johns Hopkins University, Baltimore, Maryland 21205
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Shu-Ching Huang,
Shu-Ching Huang
*Department of Medicine, The Johns Hopkins University School of Medicine, Baltimore, Maryland 21205; ‡Department of Biology, Yale University, New Haven, Connecticut 06511; §Boyer Center for Molecular Medicine and Department of Pathology, Yale School of Medicine, New Haven, Connecticut 06510; and ‖Department of Molecular Biology and Genetics, The Johns Hopkins University, Baltimore, Maryland 21205
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Julia S. Hartenstein,
Julia S. Hartenstein
*Department of Medicine, The Johns Hopkins University School of Medicine, Baltimore, Maryland 21205; ‡Department of Biology, Yale University, New Haven, Connecticut 06511; §Boyer Center for Molecular Medicine and Department of Pathology, Yale School of Medicine, New Haven, Connecticut 06510; and ‖Department of Molecular Biology and Genetics, The Johns Hopkins University, Baltimore, Maryland 21205
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Michael Snyder,
Michael Snyder
*Department of Medicine, The Johns Hopkins University School of Medicine, Baltimore, Maryland 21205; ‡Department of Biology, Yale University, New Haven, Connecticut 06511; §Boyer Center for Molecular Medicine and Department of Pathology, Yale School of Medicine, New Haven, Connecticut 06510; and ‖Department of Molecular Biology and Genetics, The Johns Hopkins University, Baltimore, Maryland 21205
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Vincent T. Marchesi,
Vincent T. Marchesi
*Department of Medicine, The Johns Hopkins University School of Medicine, Baltimore, Maryland 21205; ‡Department of Biology, Yale University, New Haven, Connecticut 06511; §Boyer Center for Molecular Medicine and Department of Pathology, Yale School of Medicine, New Haven, Connecticut 06510; and ‖Department of Molecular Biology and Genetics, The Johns Hopkins University, Baltimore, Maryland 21205
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Edward J. Benz, Jr.
Edward J. Benz, Jr.
*Department of Medicine, The Johns Hopkins University School of Medicine, Baltimore, Maryland 21205; ‡Department of Biology, Yale University, New Haven, Connecticut 06511; §Boyer Center for Molecular Medicine and Department of Pathology, Yale School of Medicine, New Haven, Connecticut 06510; and ‖Department of Molecular Biology and Genetics, The Johns Hopkins University, Baltimore, Maryland 21205
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Subhendra N. Mattagajasingh
*Department of Medicine, The Johns Hopkins University School of Medicine, Baltimore, Maryland 21205; ‡Department of Biology, Yale University, New Haven, Connecticut 06511; §Boyer Center for Molecular Medicine and Department of Pathology, Yale School of Medicine, New Haven, Connecticut 06510; and ‖Department of Molecular Biology and Genetics, The Johns Hopkins University, Baltimore, Maryland 21205
Shu-Ching Huang
*Department of Medicine, The Johns Hopkins University School of Medicine, Baltimore, Maryland 21205; ‡Department of Biology, Yale University, New Haven, Connecticut 06511; §Boyer Center for Molecular Medicine and Department of Pathology, Yale School of Medicine, New Haven, Connecticut 06510; and ‖Department of Molecular Biology and Genetics, The Johns Hopkins University, Baltimore, Maryland 21205
Julia S. Hartenstein
*Department of Medicine, The Johns Hopkins University School of Medicine, Baltimore, Maryland 21205; ‡Department of Biology, Yale University, New Haven, Connecticut 06511; §Boyer Center for Molecular Medicine and Department of Pathology, Yale School of Medicine, New Haven, Connecticut 06510; and ‖Department of Molecular Biology and Genetics, The Johns Hopkins University, Baltimore, Maryland 21205
Michael Snyder
*Department of Medicine, The Johns Hopkins University School of Medicine, Baltimore, Maryland 21205; ‡Department of Biology, Yale University, New Haven, Connecticut 06511; §Boyer Center for Molecular Medicine and Department of Pathology, Yale School of Medicine, New Haven, Connecticut 06510; and ‖Department of Molecular Biology and Genetics, The Johns Hopkins University, Baltimore, Maryland 21205
Vincent T. Marchesi
*Department of Medicine, The Johns Hopkins University School of Medicine, Baltimore, Maryland 21205; ‡Department of Biology, Yale University, New Haven, Connecticut 06511; §Boyer Center for Molecular Medicine and Department of Pathology, Yale School of Medicine, New Haven, Connecticut 06510; and ‖Department of Molecular Biology and Genetics, The Johns Hopkins University, Baltimore, Maryland 21205
Edward J. Benz, Jr.
*Department of Medicine, The Johns Hopkins University School of Medicine, Baltimore, Maryland 21205; ‡Department of Biology, Yale University, New Haven, Connecticut 06511; §Boyer Center for Molecular Medicine and Department of Pathology, Yale School of Medicine, New Haven, Connecticut 06510; and ‖Department of Molecular Biology and Genetics, The Johns Hopkins University, Baltimore, Maryland 21205
Address correspondence to Shu-Ching Huang, 1033 Ross Research Building, 720 Rutland Avenue, Department of Medicine, Johns Hopkins University, Baltimore, MD 21205. Tel.: 410-614-4610. Fax: 410-955-0185. E-mail: [email protected]
Received:
May 29 1998
Revision Received:
February 26 1999
Online ISSN: 1540-8140
Print ISSN: 0021-9525
1999
J Cell Biol (1999) 145 (1): 29–43.
Article history
Received:
May 29 1998
Revision Received:
February 26 1999
Citation
Subhendra N. Mattagajasingh, Shu-Ching Huang, Julia S. Hartenstein, Michael Snyder, Vincent T. Marchesi, Edward J. Benz; A Nonerythroid Isoform of Protein 4.1R Interacts with the Nuclear Mitotic Apparatus (NuMA) Protein . J Cell Biol 5 April 1999; 145 (1): 29–43. doi: https://doi.org/10.1083/jcb.145.1.29
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