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A genomic toolkit to investigate kinesin and myosin motor function in cells
- Source :
- Nature cell biology. 15(3)
- Publication Year :
- 2012
-
Abstract
- Coordination of multiple kinesin and myosin motors is required for intracellular transport, cell motility and mitosis. However, comprehensive resources that allow systems analysis of the localization and interplay between motors in living cells do not exist. Here, we generated a library of 243 amino- and carboxy-terminally tagged mouse and human bacterial artificial chromosome transgenes to establish 227 stably transfected HeLa cell lines, 15 mouse embryonic stem cell lines and 1 transgenic mouse line. The cells were characterized by expression and localization analyses and further investigated by affinity-purification mass spectrometry, identifying 191 candidate protein-protein interactions. We illustrate the power of this resource in two ways. First, by characterizing a network of interactions that targets CEP170 to centrosomes, and second, by showing that kinesin light-chain heterodimers bind conventional kinesin in cells. Our work provides a set of validated resources and candidate molecular pathways to investigate motor protein function across cell lineages.
- Subjects :
- Chromosomes, Artificial, Bacterial
Fluorescent Antibody Technique
Kinesins
Microtubules
Chromatography, Affinity
Mice
Neuroblastoma
0302 clinical medicine
Cell Movement
Myosin
Transgenes
Phylogeny
Oligonucleotide Array Sequence Analysis
Neurons
0303 health sciences
Reverse Transcriptase Polymerase Chain Reaction
Stem Cells
Transfection
Genomics
Cell biology
Kinesin
Microtubule-Associated Proteins
Blotting, Western
Green Fluorescent Proteins
Mitosis
Mice, Transgenic
Biology
Myosins
Real-Time Polymerase Chain Reaction
Motor protein
03 medical and health sciences
Microtubule
Animals
Humans
Immunoprecipitation
RNA, Messenger
Embryonic Stem Cells
030304 developmental biology
Centrosome
Gene Expression Profiling
Biological Transport
Cell Biology
Phosphoproteins
Cell culture
Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
Protein Multimerization
030217 neurology & neurosurgery
Biomarkers
HeLa Cells
Subjects
Details
- ISSN :
- 14764679
- Volume :
- 15
- Issue :
- 3
- Database :
- OpenAIRE
- Journal :
- Nature cell biology
- Accession number :
- edsair.doi.dedup.....4776e732a023823c343f6db27b2eb7ec