that ulk3 phosphorylates gli proteins in vitro and gli1 has at least two phosphorylation sites situated in n and c terminus of the protein we have found that in contrast to stk36 the kinase deficient mutants of ulk3 are inactive indicating that functional kinase domain of ulk3 is required for the regulation of gli protein activity also we show that ulk3 expression is higher in fetal brain and in a number of postnatal tissues where shh signaling is known to be active our data suggests that ulk3 is involved in shh pathway as a positive regulator of gli proteins in this disclosure we identify human ulk3 as a serine threonine kinase regulating positively shh pathway in mammalian cells it has been shown previously that protein kinase dfu is absolutely required for the activation of hh pathway in drosophila dfu works in a concert with cos2 dsmo and dsufu forming a microtubule binding complex that controls activity of ci transcription factor 18 20 22 23 in spite of overall hh pathway conservation between vertebrates and invertebrates the function of those proteins is not fully conserved in mammals in drosophila cos2 plays a central role in regulating the activity of ci 40 41 but mammalian orthologues of cos2 kif7 and kif27 have no effect on shh signaling 4 the cytoplasmic c terminus of dsmo that is extremely important for hh signaling in.