Document Detail


Multiple localization of endogenous MARK4L protein in human glioma.
MedLine Citation:
PMID:  19759416     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
BACKGROUND: We have previously shown that the sustained expression of MARK4L transcripts in glioma and neural progenitors (NHNPs) declines after exposure to antisense MARK4L oligonucleotides in glioblastoma cell lines. Array-CGH confirmed the genomic duplication of MARK4L identified by FISH in a glioblastoma cell line. This background together with literature data on the exogenous association of MARK4 with interphase centrosome prompted us to investigate the sub-cellular localization of the endogenous MARK4L protein aiming at achieving insights on its possible role in the pathomechanisms of glioma. METHODS: Immunodetection was carried out to validate the specificity of MARK4L antibody in gliomas and NHNPs. Mass spectrometry was applied for MARK4L protein identification in a representative glioblastoma cell line. Combined biochemical fractionation and immunodetection analyses were performed to confirm the sub-cellular localization of MARK4L achieved by immunofluorescence in glioma cell lines. RESULTS: By assigning MARK4L protein within the band immunoprecipitated by the specific antibody we validated our anti-MARK4L antibody. We demonstrated that the endogenous MARK4L: (i) colocalizes with centrosomes at all mitotic stages and resides in centrosome-enriched fractions; (ii) associates with the nucleolus and the midbody and respective fractions, and (iii) co-stains the aberrant centrosome configurations observed in glioma cell lines. CONCLUSION: The overall data merge on the multiplex entry of MARK4L into the cell cycle and link it to the aberrant centrosomes in glioma cell lines suggesting a possible role of this kinase in the abnormal mitotic processes of human glioma.
Authors:
Ivana Magnani; Chiara Novielli; Melissa Bellini; Gaia Roversi; Lorenzo Bello; Lidia Larizza
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't    
Journal Detail:
Title:  Cellular oncology : the official journal of the International Society for Cellular Oncology     Volume:  31     ISSN:  1875-8606     ISO Abbreviation:  Cell. Oncol.     Publication Date:  2009  
Date Detail:
Created Date:  2009-09-17     Completed Date:  2009-12-21     Revised Date:  -    
Medline Journal Info:
Nlm Unique ID:  101219418     Medline TA:  Cell Oncol     Country:  Netherlands    
Other Details:
Languages:  eng     Pagination:  357-70     Citation Subset:  IM    
Affiliation:
Dipartimento di Medicina, Chirurgia e Odontoiatria, Sezione di Genetica Medica, Universit?? di Milano, Milan, Italy.
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MeSH Terms
Descriptor/Qualifier:
Antibodies / immunology
Cell Line, Tumor
Cell Nucleus / metabolism
Centrosome / metabolism
Glioma / metabolism*,  pathology
Humans
Immunoprecipitation
Interphase
Mass Spectrometry
Mitosis
Molecular Weight
Neurons / metabolism
Protein-Serine-Threonine Kinases / chemistry,  immunology,  metabolism*
Stem Cells / metabolism
Subcellular Fractions / metabolism
Chemical
Reg. No./Substance:
0/Antibodies; EC 2.7.1.-/MARK4 protein, human; EC 2.7.11.1/Protein-Serine-Threonine Kinases

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