The NH2-terminal region of the active domain of sonic hedgehog is necessary for its signal transduction.
The NH2-terminal domain of sonic hedgehog (residue 25-198) was expressed in both yeast and animal cells. The yeast-derived NH2-terminal domain of sonic hedgehog was less active by far than the animal cell-derived counterpart. The yeast-derived NH2-terminal domain of sonic hedgehog lacked 10 amino acids from the NH2-terminus. This cleavage of the yeast-derived NH2-terminal domain of sonic hedgehog might due to Kex 2. In contrast, a mutant yeast-derived NH2-terminal domain of sonic hedgehog (Lys-33 to Thr) retained its NH2-terminus and its activity was comparable to that of the animal cell-derived NH2-terminal domain of sonic hedgehog. The NH2-terminal deleted NH2-terminal domain of sonic hedgehog completely lost its activity, nevertheless it inhibited the alkaline phosphatase activity induced by the animal cell-derived NH2-terminal domain of sonic hedgehog in a dose-dependent manner. These data suggest that the NH2-terminal deleted NH2-terminal domain of sonic hedgehog retains a receptor-binding ability and that the NH2-terminal peptide of the NH2-terminal domain of sonic hedgehog is necessary for its signal transduction.
Katsuura
M
M
Discovery Research Laboratories, Hoechst Marion Roussel, Kawagoe, Saitama, Japan. m.katsuura@teijin.co.jp
Hosono-Sakuma
Y
Y
Wagatsuma
M
M
Yanagisawa
S
S
Okazaki
M
M
Kimura
M
M
D000469:Alkaline Phosphatase / Q000037:antagonists & inhibitors; D019943:Amino Acid Substitution; D000818:Animals; D001483:Base Sequence; D001665:Binding Sites / Q000235:genetics; D017931:DNA Primers / Q000235:genetics; D015870:Gene Expression; D053823:Hedgehog Proteins; D066298:In Vitro Techniques; D007739:L Cells; D051379:Mice; D011506:Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D012441:Saccharomyces cerevisiae / Q000235:genetics; D017384:Sequence Deletion; D015398:Signal Transduction; D015534:Trans-Activators
D017931:DNA Primers; D053823:Hedgehog Proteins; D011506:Proteins; D011994:Recombinant Proteins; D015534:Trans-Activators; D000469:Alkaline Phosphatase