Document Detail

The mitotic arrest deficient protein MAD2B interacts with the clathrin light chain A during mitosis.
MedLine Citation:
PMID:  21152103     Owner:  NLM     Status:  MEDLINE    
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.
Klaas Medendorp; Lilian Vreede; Jan J M van Groningen; Lisette Hetterschijt; Linda Brugmans; Patrick A M Jansen; Wilhelmina H van den Hurk; Diederik R H de Bruijn; Ad Geurts van Kessel
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't     Date:  2010-11-30
Journal Detail:
Title:  PloS one     Volume:  5     ISSN:  1932-6203     ISO Abbreviation:  PLoS ONE     Publication Date:  2010  
Date Detail:
Created Date:  2010-12-14     Completed Date:  2011-04-27     Revised Date:  2013-07-03    
Medline Journal Info:
Nlm Unique ID:  101285081     Medline TA:  PLoS One     Country:  United States    
Other Details:
Languages:  eng     Pagination:  e15128     Citation Subset:  IM    
Department of Human Genetics, Radboud University Nijmegen Medical Centre, Nijmegen Centre for Molecular Life Sciences, Nijmegen, The Netherlands.
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MeSH Terms
Basic Helix-Loop-Helix Leucine Zipper Transcription Factors / genetics,  metabolism
Blotting, Western
Cell Cycle Proteins / genetics,  metabolism
Cell Line, Tumor
Clathrin Light Chains / genetics,  metabolism*
G2 Phase
HEK293 Cells
Luminescent Proteins / genetics,  metabolism
Microscopy, Confocal
Mitotic Spindle Apparatus / metabolism*
Neoplasm Proteins / genetics,  metabolism
Protein Binding
Proteins / genetics,  metabolism*
RNA Interference
Recombinant Fusion Proteins / genetics,  metabolism
Reg. No./Substance:
0/Basic Helix-Loop-Helix Leucine Zipper Transcription Factors; 0/Cell Cycle Proteins; 0/Clathrin Light Chains; 0/Luminescent Proteins; 0/MAD2L2 protein, human; 0/Neoplasm Proteins; 0/PRCC protein, human; 0/Proteins; 0/Recombinant Fusion Proteins; 0/TFE3 protein, human

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine

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