Dorina B. Wolf, Dieter Maier, and Anja C. Nagel
Department of General Genetics (190g), University of Hohenheim, Garbenstr. 30, 70599 Stuttgart, Germany
CSL transcription factors are central to signal transduction in the highly conserved Notch signaling pathway. CSL acts as a molecular switch: depending on the cofactors recruited, CSL induces either activation or repression of Notch target genes. Unexpectedly, CSL depends on its cofactors for nuclear entry, despite its role as gene regulator. In Drosophila, the CSL homologue Suppressor of Hairless (Su(H)), recruits Hairless (H) for repressor complex assembly, and eventually for nuclear import. We recently found that Su(H) is subjected to a dynamic nucleo-cytoplasmic shuttling, thereby strictly following H subcellular distribution. Hence, regulation of nuclear availability of Su(H) by H may represent a new layer of control of Notch signaling activity. Here we extended this work on the murine CSL homologue RBPJ. Using a ‘murinized’ fly model bearing RBPJwt in place of Su(H) at the endogenous locus we demonstrate that RBPJ protein likewise follows H subcellular distribution. For example, overexpression of a H*NLS3 protein variant defective of nuclear import resulted in a cytosolic localization of RBPJ protein, whereas the overexpression of a H*NES protein variant defective in the nuclear export signal caused the accumulation of RBPJ protein in the nucleus. Evidently, RBPJ is exported from the nucleus as well. Overall these data demonstrate that in our fly model, RBPJ is subjected to H-mediated nucleo-cytoplasmic shuttling as is Su(H). These data raise the possibility that nuclear availability of mammalian CSL proteins is likewise restricted by cofactors, and may hence present a more general mode of regulating Notch signaling activity.
A Leica MZ FLIII UV stereo-microscope (Leica, Wetzlar, Germany) illuminated with CoolLED pE-300white (AHF, Tübingen, Germany) was used for the purpose of selecting the larvae.
Product Associated Features
The pE-300white is a popular illuminator for fluorescent screening of Drosophila larvae, with individual control of three LED channels. The blue LED channel can be used individually to enhance the GFP contrast, and the irradiance can be easily balanced with the control pod for optimum illumination.
Year of Publication
Country of Publication