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Internalization and Down-Regulation of the ALK Receptor in Neuroblastoma Cell Lines upon Monoclonal Antibodies Treatment
- Source :
- PLoS ONE, PLoS ONE, Vol 7, Iss 3, p e33581 (2012)
- Publication Year :
- 2012
- Publisher :
- Public Library of Science (PLoS), 2012.
-
Abstract
- Recently, activating mutations of the full length ALK receptor, with two hot spots at positions F1174 and R1275, have been characterized in sporadic cases of neuroblastoma. Here, we report similar basal patterns of ALK phosphorylation between the neuroblastoma IMR-32 cell line, which expresses only the wild-type receptor (ALK(WT)), and the SH-SY5Y cell line, which exhibits a heterozygous ALK F1174L mutation and expresses both ALK(WT) and ALK(F1174L) receptors. We demonstrate that this lack of detectable increased phosphorylation in SH-SY5Y cells is a result of intracellular retention and proteasomal degradation of the mutated receptor. As a consequence, in SH-SY5Y cells, plasma membrane appears strongly enriched for ALK(WT) whereas both ALK(WT) and ALK(F1174L) were present in intracellular compartments. We further explored ALK receptor trafficking by investigating the effect of agonist and antagonist mAb (monoclonal antibodies) on ALK internalization and down-regulation, either in SH-SY5Y cells or in cells expressing only ALK(WT). We observe that treatment with agonist mAbs resulted in ALK internalization and lysosomal targeting for receptor degradation. In contrast, antagonist mAb induced ALK internalization and recycling to the plasma membrane. Importantly, we correlate this differential trafficking of ALK in response to mAb with the recruitment of the ubiquitin ligase Cbl and ALK ubiquitylation only after agonist stimulation. This study provides novel insights into the mechanisms regulating ALK trafficking and degradation, showing that various ALK receptor pools are regulated by proteasome or lysosome pathways according to their intracellular localization.
- Subjects :
- Transcription, Genetic
Cell- och molekylärbiologi
Receptor Protein-Tyrosine Kinases
lcsh:Medicine
Biochemistry
Neuroblastoma
Nucleic Acids
hemic and lymphatic diseases
Molecular Cell Biology
Anaplastic lymphoma kinase
Anaplastic Lymphoma Kinase
Phosphorylation
lcsh:Science
Internalization
Receptor
media_common
Multidisciplinary
biology
Antibodies, Monoclonal
Signaling Cascades
Ubiquitin ligase
Cell biology
Protein Transport
medicine.anatomical_structure
Oncology
Medicine
Intracellular
Research Article
Signal Transduction
Agonist
Proteasome Endopeptidase Complex
medicine.drug_class
media_common.quotation_subject
Down-Regulation
Cell Line, Tumor
Lysosome
medicine
Humans
Biology
Base Sequence
lcsh:R
Cancers and Neoplasms
DNA
Molecular biology
Enzyme Activation
Mutation
biology.protein
lcsh:Q
Cell and Molecular Biology
Subjects
Details
- ISSN :
- 19326203
- Volume :
- 7
- Database :
- OpenAIRE
- Journal :
- PLoS ONE
- Accession number :
- edsair.doi.dedup.....18f900a8062559e6127dfce8bc562a74