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. Author manuscript; available in PMC: 2014 Jul 1.

Published in final edited form as: Nat Commun. 2014;5:3318. doi: 10.1038/ncomms4318

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S109 phosphorylation of ARPP19 prevents the initial activation of Cdk1 required for meiosis resumption.

(a) Injection of S109D-GST-ARPP inhibits Cdk1 activation induced either by PKI or Cyclin B. Prophase-arrested oocytes (Pro) were injected with 300 ng of WT- or S109D-GST-ARPP then induced to mature by Pg or by injecting either a non-degradable form of Cyclin B (CycB) or PKI. Oocytes were collected at GVBD and Cdk1 activation was followed by immunoblotting Gwl, Cyclin B2 (CycB2), MAPK phosphorylation (pMAPK), Cdk1 phosphorylation at Y15 (pY15-Cdk1) and phosphorylated Cdk1 substrates (pCdk Sub.).

(b) Injection of S109D-GST-ARPP prevents Mos-induced Cdk1 activation but does not inhibit meiotic resumption induced by S67-thiophosphorylated WT-GST-ARPP. Prophase-arrested oocytes (Pro) were injected or not with 300 ng of S109D-GST-ARPP (S109D) then induced to mature by progesterone (Pg) or by injecting either Mos or S67-thiophosphorylated WT-GST-ARPP (WT*). Oocytes were collected at GVBD and immunoblotted for Gwl, Cyclin B2 upshift (CycB2), Cdk1 phosphorylation at Y15 (pY15-Cdk1), phosphorylated MAPK (pMAPK) and phosphorylated Cdk1 substrates (pCdk Sub.).

(c) Injection of S109D-GST-ARPP does not inhibit Cdk1 activation induced by K71M-Gwl. Prophase-arrested oocytes (Pro) were injected with WT-, S67A-, S109A- or S109D-GST-ARPP and meiotic maturation was triggered by the injection of mRNA encoding K71M-Gwl. Oocytes were collected 18 hours later and immunoblotted for phosphorylated Cdc27, Mos accumulation and phosphorylated MAPK (pMAPK).