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The Mitotic Arrest Deficient Protein MAD2B Interacts with the Clathrin Light Chain A during Mitosis

BACKGROUND: Although the mitotic arrest deficient protein MAD2B (MAD2L2) is thought to inhibit the anaphase promoting complex (APC) by binding to CDC20 and/or CDH1 (FZR1), its exact role in cell cycle control still remains to be established. METHODOLOGY/PRINCIPAL FINDINGS: Using a yeast two-hybrid i...

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Autores principales: Medendorp, Klaas, Vreede, Lilian, van Groningen, Jan J. M., Hetterschijt, Lisette, Brugmans, Linda, Jansen, Patrick A. M., van den Hurk, Wilhelmina H., de Bruijn, Diederik R. H., van Kessel, Ad Geurts
Formato: Texto
Lenguaje:English
Publicado: Public Library of Science 2010
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2994903/
https://www.ncbi.nlm.nih.gov/pubmed/21152103
http://dx.doi.org/10.1371/journal.pone.0015128
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author Medendorp, Klaas
Vreede, Lilian
van Groningen, Jan J. M.
Hetterschijt, Lisette
Brugmans, Linda
Jansen, Patrick A. M.
van den Hurk, Wilhelmina H.
de Bruijn, Diederik R. H.
van Kessel, Ad Geurts
author_facet Medendorp, Klaas
Vreede, Lilian
van Groningen, Jan J. M.
Hetterschijt, Lisette
Brugmans, Linda
Jansen, Patrick A. M.
van den Hurk, Wilhelmina H.
de Bruijn, Diederik R. H.
van Kessel, Ad Geurts
author_sort Medendorp, Klaas
collection PubMed
description BACKGROUND: Although the mitotic arrest deficient protein MAD2B (MAD2L2) is thought to inhibit the anaphase promoting complex (APC) by binding to CDC20 and/or CDH1 (FZR1), its exact role in cell cycle control still remains to be established. METHODOLOGY/PRINCIPAL FINDINGS: Using a yeast two-hybrid interaction trap we identified the human clathrin light chain A (CLTA) as a novel MAD2B binding protein. A direct interaction was established in mammalian cells via GST pull-down and endogenous co-immunoprecipitation during the G2/M phase of the cell cycle. Through subsequent confocal laser scanning microscopy we found that MAD2B and CLTA co-localize at the mitotic spindle. Clathrin forms a trimeric structure, i.e., the clathrin triskelion, consisting of three heavy chains (CLTC), each with an associated light chain. This clathrin structure has previously been shown to be required for the function of the mitotic spindle through stabilization of kinetochore fibers. Upon siRNA-mediated MAD2B depletion, we found that CLTA was no longer concentrated at the mitotic spindle but, instead, diffusely distributed throughout the cell. In addition, we found a marked increase in the percentage of misaligned chromosomes. CONCLUSIONS/SIGNIFICANCE: Previously, we identified MAD2B as an interactor of the renal cell carcinoma (RCC)-associated protein PRCC. In addition, we found that fusion of PRCC with the transcription factor TFE3 in t(X;1)(p11;q21)-positive RCCs results in an impairment of this interaction and a concomitant failure to shuttle MAD2B to the nucleus. Our current data show that MAD2B interacts with CLTA during the G2/M phase of the cell cycle and that depletion of MAD2B leads to a marked increase in the percentage of misaligned chromosomes and a redistribution of CLTA during mitosis.
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spelling pubmed-29949032010-12-10 The Mitotic Arrest Deficient Protein MAD2B Interacts with the Clathrin Light Chain A during Mitosis Medendorp, Klaas Vreede, Lilian van Groningen, Jan J. M. Hetterschijt, Lisette Brugmans, Linda Jansen, Patrick A. M. van den Hurk, Wilhelmina H. de Bruijn, Diederik R. H. van Kessel, Ad Geurts PLoS One Research Article BACKGROUND: Although the mitotic arrest deficient protein MAD2B (MAD2L2) is thought to inhibit the anaphase promoting complex (APC) by binding to CDC20 and/or CDH1 (FZR1), its exact role in cell cycle control still remains to be established. METHODOLOGY/PRINCIPAL FINDINGS: Using a yeast two-hybrid interaction trap we identified the human clathrin light chain A (CLTA) as a novel MAD2B binding protein. A direct interaction was established in mammalian cells via GST pull-down and endogenous co-immunoprecipitation during the G2/M phase of the cell cycle. Through subsequent confocal laser scanning microscopy we found that MAD2B and CLTA co-localize at the mitotic spindle. Clathrin forms a trimeric structure, i.e., the clathrin triskelion, consisting of three heavy chains (CLTC), each with an associated light chain. This clathrin structure has previously been shown to be required for the function of the mitotic spindle through stabilization of kinetochore fibers. Upon siRNA-mediated MAD2B depletion, we found that CLTA was no longer concentrated at the mitotic spindle but, instead, diffusely distributed throughout the cell. In addition, we found a marked increase in the percentage of misaligned chromosomes. CONCLUSIONS/SIGNIFICANCE: Previously, we identified MAD2B as an interactor of the renal cell carcinoma (RCC)-associated protein PRCC. In addition, we found that fusion of PRCC with the transcription factor TFE3 in t(X;1)(p11;q21)-positive RCCs results in an impairment of this interaction and a concomitant failure to shuttle MAD2B to the nucleus. Our current data show that MAD2B interacts with CLTA during the G2/M phase of the cell cycle and that depletion of MAD2B leads to a marked increase in the percentage of misaligned chromosomes and a redistribution of CLTA during mitosis. Public Library of Science 2010-11-30 /pmc/articles/PMC2994903/ /pubmed/21152103 http://dx.doi.org/10.1371/journal.pone.0015128 Text en Medendorp et al. http://creativecommons.org/licenses/by/4.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are properly credited.
spellingShingle Research Article
Medendorp, Klaas
Vreede, Lilian
van Groningen, Jan J. M.
Hetterschijt, Lisette
Brugmans, Linda
Jansen, Patrick A. M.
van den Hurk, Wilhelmina H.
de Bruijn, Diederik R. H.
van Kessel, Ad Geurts
The Mitotic Arrest Deficient Protein MAD2B Interacts with the Clathrin Light Chain A during Mitosis
title The Mitotic Arrest Deficient Protein MAD2B Interacts with the Clathrin Light Chain A during Mitosis
title_full The Mitotic Arrest Deficient Protein MAD2B Interacts with the Clathrin Light Chain A during Mitosis
title_fullStr The Mitotic Arrest Deficient Protein MAD2B Interacts with the Clathrin Light Chain A during Mitosis
title_full_unstemmed The Mitotic Arrest Deficient Protein MAD2B Interacts with the Clathrin Light Chain A during Mitosis
title_short The Mitotic Arrest Deficient Protein MAD2B Interacts with the Clathrin Light Chain A during Mitosis
title_sort mitotic arrest deficient protein mad2b interacts with the clathrin light chain a during mitosis
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2994903/
https://www.ncbi.nlm.nih.gov/pubmed/21152103
http://dx.doi.org/10.1371/journal.pone.0015128
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