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Title: Antibody-based analysis reveals “filamentous vs. non-filamentous” and “cytoplasmic vs. nuclear” crosstalk of cytoskeletal proteins

Abstract

To uncover the molecular composition and dynamics of the functional scaffold for the nucleus, three fractions of biochemically-stable nuclear protein complexes were extracted and used as immunogens to produce a variety of monoclonal antibodies. Many helix-based cytoskeletal proteins were identified as antigens, suggesting their dynamic contribution to nuclear architecture and function. Interestingly, sets of antibodies distinguished distinct subcellular localization of a single isoform of certain cytoskeletal proteins; distinct molecular forms of keratin and actinin were found in the nucleus. Their nuclear shuttling properties were verified by the apparent nuclear accumulations under inhibition of CRM1-dependent nuclear export. Nuclear keratins do not take an obvious filamentous structure, as was revealed by non-filamentous cytoplasmic keratin-specific monoclonal antibody. These results suggest the distinct roles of the helix-based cytoskeletal proteins in the nucleus. - Highlights: • A set of monoclonal antibodies were raised against nuclear scaffold proteins. • Helix-based cytoskeletal proteins were involved in nuclear scaffold. • Many cytoskeletal components shuttle into the nucleus in a CRM1-dependent manner. • Sets of antibodies distinguished distinct subcellular localization of a single isoform. • Nuclear keratin is soluble and does not form an obvious filamentous structure.

Authors:
 [1]; ;  [1];  [2];  [1]
  1. Graduate School of Biostudies, Kyoto University, Kyoto 606-8501 (Japan)
  2. Graduate School of Science and Technology, Niigata University, Niigata 950-2181 (Japan)
Publication Date:
OSTI Identifier:
22278205
Resource Type:
Journal Article
Resource Relation:
Journal Name: Experimental Cell Research; Journal Volume: 319; Journal Issue: 20; Other Information: Copyright (c) 2013 Elsevier Science B.V., Amsterdam, The Netherlands, All rights reserved.; Country of input: International Atomic Energy Agency (IAEA)
Country of Publication:
United States
Language:
English
Subject:
60 APPLIED LIFE SCIENCES; ANTIGENS; KERATIN; MICROTUBULES; MONOCLONAL ANTIBODIES

Citation Formats

Kumeta, Masahiro, E-mail: kumeta@lif.kyoto-u.ac.jp, Hirai, Yuya, Yoshimura, Shige H., Horigome, Tsuneyoshi, and Takeyasu, Kunio. Antibody-based analysis reveals “filamentous vs. non-filamentous” and “cytoplasmic vs. nuclear” crosstalk of cytoskeletal proteins. United States: N. p., 2013. Web. doi:10.1016/J.YEXCR.2013.07.021.
Kumeta, Masahiro, E-mail: kumeta@lif.kyoto-u.ac.jp, Hirai, Yuya, Yoshimura, Shige H., Horigome, Tsuneyoshi, & Takeyasu, Kunio. Antibody-based analysis reveals “filamentous vs. non-filamentous” and “cytoplasmic vs. nuclear” crosstalk of cytoskeletal proteins. United States. doi:10.1016/J.YEXCR.2013.07.021.
Kumeta, Masahiro, E-mail: kumeta@lif.kyoto-u.ac.jp, Hirai, Yuya, Yoshimura, Shige H., Horigome, Tsuneyoshi, and Takeyasu, Kunio. Tue . "Antibody-based analysis reveals “filamentous vs. non-filamentous” and “cytoplasmic vs. nuclear” crosstalk of cytoskeletal proteins". United States. doi:10.1016/J.YEXCR.2013.07.021.
@article{osti_22278205,
title = {Antibody-based analysis reveals “filamentous vs. non-filamentous” and “cytoplasmic vs. nuclear” crosstalk of cytoskeletal proteins},
author = {Kumeta, Masahiro, E-mail: kumeta@lif.kyoto-u.ac.jp and Hirai, Yuya and Yoshimura, Shige H. and Horigome, Tsuneyoshi and Takeyasu, Kunio},
abstractNote = {To uncover the molecular composition and dynamics of the functional scaffold for the nucleus, three fractions of biochemically-stable nuclear protein complexes were extracted and used as immunogens to produce a variety of monoclonal antibodies. Many helix-based cytoskeletal proteins were identified as antigens, suggesting their dynamic contribution to nuclear architecture and function. Interestingly, sets of antibodies distinguished distinct subcellular localization of a single isoform of certain cytoskeletal proteins; distinct molecular forms of keratin and actinin were found in the nucleus. Their nuclear shuttling properties were verified by the apparent nuclear accumulations under inhibition of CRM1-dependent nuclear export. Nuclear keratins do not take an obvious filamentous structure, as was revealed by non-filamentous cytoplasmic keratin-specific monoclonal antibody. These results suggest the distinct roles of the helix-based cytoskeletal proteins in the nucleus. - Highlights: • A set of monoclonal antibodies were raised against nuclear scaffold proteins. • Helix-based cytoskeletal proteins were involved in nuclear scaffold. • Many cytoskeletal components shuttle into the nucleus in a CRM1-dependent manner. • Sets of antibodies distinguished distinct subcellular localization of a single isoform. • Nuclear keratin is soluble and does not form an obvious filamentous structure.},
doi = {10.1016/J.YEXCR.2013.07.021},
journal = {Experimental Cell Research},
number = 20,
volume = 319,
place = {United States},
year = {Tue Dec 10 00:00:00 EST 2013},
month = {Tue Dec 10 00:00:00 EST 2013}
}