1. Regulation of RECQ4–MCM10 interaction by CDK phosphorylation.
Regulation of RECQ4–MCM10 interaction by CDK phosphorylation. (A) Phosphorylation of GST–RECQ41−200 by 293T extracts. Glutathione beads with GST–RECQ41−200 fragment were incubated with asynchronized 293T extracts without (left) or with (right) roscovitine in the presence of γ32P‐ATP. After washing, 32P‐labelled products were detected by SDS–PAGE followed by Coomassie blue staining and autoradiography. (B) Sequence of the first 200a.a. of RECQ4 containing the Sld2 domain. The light grey boxes indicate the cyclin recognition motif (RxL); the dark grey boxes indicate the potential CDK target sites. (C) Phosphorylation of GST–RECQ41−200 WT or glutamate mutants by 293T extracts as described in (A). (D) GST–MCM10 pull‐down of full‐length WT RECQ4 or glutamate substituted triple mutant at S89, T93 and T139. Inputs and bound fractions were analysed on SDS–PAGE followed by western blotting using anti‐His antibody.
Xiaohua Xu et al. EMBO J. 2009;28:
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