Examination
of keratin filament network biogenesis and dynamics in murine embryos
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| Keratins
form the major cytoskeleton of epithelial cells and behave at the same
time as a highly dynamic protein scaffold. As such, they protect
epithelia against mechanical stress but also play a major role in the
regulation of cell growth, proliferation, stress pathways and organelle
transport. The molecular mechanisms governing the localized assembly of
keratins from heterodimeric subunits and their interaction with cell
adhesion molecules and regulatory proteins are not well understood but
are essential for an understanding of keratinopathies. Based on high
resolution live cell imaging of cell transfectants we have recently
developed a model for keratin assembly and the involvement of p38 MAP
kinases that allows predictions on the in vivo properties of the
keratin cytoskeleton. To evaluate these hypotheses, we propose to
generate a knock-in allele coding for a hybrid fluorescent keratin 8.
This will enable us to monitor keratin assembly and its MAP
kinase-dependent regulation by direct fluorescence microscopy in
pre- and postimplantation embryos, and in the long run, in postnatal
mice. The contribution of other cytoskeletal components, desmosomal
proteins and of certain kinases will be studied by genetic and
pharmacological approaches in embryonal stem cell derivatives and mouse
embryos. As a future perspective, fluorescent keratin 8 knock-in
animals will be employed to elucidate the interdependence of desmosomes
and adherens junctions with keratins, using mouse strains deficient in
the relevant genes.
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Principal
investigator: Dr. Anne Kölsch
Funding: START
Nachwuchsprogramm
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