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Multiple Localization of Endogenous MARK4L Protein in Human Glioma
- Source :
- Cellular Oncology, Vol 31, Iss 5, Pp 357-370 (2009), Cellular Oncology : the Official Journal of the International Society for Cellular Oncology
- Publication Year :
- 2009
- Publisher :
- Hindawi Limited, 2009.
-
Abstract
- 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.Conclusions: 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.
- Subjects :
- Cancer Research
Antibodie
Mitosis
Protein Serine-Threonine Kinases
Protein-Serine-Threonine Kinase
lcsh:RC254-282
Antibodies
Mass Spectrometry
Pathology and Forensic Medicine
Stem Cell
Cell Line, Tumor
Humans
MARK4L
Immunoprecipitation
centrosome abnormalities
lcsh:QH573-671
Cell Nucleu
Interphase
neoplasms
Cell Nucleus
Neurons
Centrosome
lcsh:Cytology
Stem Cells
Cell Biology
General Medicine
Glioma
Neuron
Mitosi
lcsh:Neoplasms. Tumors. Oncology. Including cancer and carcinogens
nervous system diseases
Molecular Weight
immunodetection
multiple subcellular localization
Molecular Medicine
Other
Human
Subcellular Fractions
Subjects
Details
- Language :
- English
- ISSN :
- 18758606 and 15705870
- Volume :
- 31
- Issue :
- 5
- Database :
- OpenAIRE
- Journal :
- Cellular Oncology
- Accession number :
- edsair.doi.dedup.....dab0da07d140979222caacc78e126509