Botch Is a γ-Glutamyl Cyclotransferase that Deglycinates and Antagonizes Notch

•Botch has γ-glutamyl cyclotransferase activity•Notch is monoglycinated on the γ-glutamyl carbon of glutamate 1,669•Botch deglycinates Notch, preventing S1 furin-like cleavage and thus Notch signaling•Botch creates a 5-oxy-proline posttranslational modification at Notch glutamate 1,669

SummaryBotch promotes embryonic neurogenesis by inhibiting the initial S1 furin-like cleavage step of Notch maturation. The biochemical process by which Botch inhibits Notch maturation is not known. Here, we show that Botch has γ-glutamyl cyclotransferase (GGCT) activity that deglycinates Notch, which prevents the S1 furin-like cleavage. Moreover, Notch is monoglycinated on the γ-glutamyl carbon of glutamate 1,669. The deglycinase activity of Botch is required for inhibition of Notch signaling both in vitro and in vivo. When the γ-glutamyl-glycine at position 1,669 of Notch is degylcinated, it is replaced by 5-oxy-proline. These results reveal that Botch regulates Notch signaling through deglycination and identify a posttranslational modification of Notch that plays an important role in neurogenesis.

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