ULK1 is a serine/threonine protein kinase that plays critical role during initial stages of autophagy which is a vital response to nutrient starvation. The conserved C-terminal domain (CTD) of ULK1 controls the regulatory function and localization of the protein. Knockdown of ULK1 inhibits the autophagic response as well as inhibiting rapamycin-induced autophagy consistent with a role downstream of mTOR. ULK1 forms a complex with FIP200 and ATG13 and this complex is essential for starvation-induced autophagy. Both FIP200 and ATG13 are critical for correct localization of ULK1 to the pre-autophagosome and stability of ULK1 protein. ULK1 is phosphorylated by the mTOR pathway in a nutrient starvation-regulated manner. Recombinant human ULK1 (1-649) was expressed by baculovirus in Sf9 insect cells using an N-terminal GST tag.
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