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. 2005 Oct;16(10):5026–5039. doi: 10.1091/mbc.E05-05-0412

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Copyright © 2005, The American Society for Cell Biology

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Figure 8. — The Mcm3 consensus CDK phosphorylation sites are required for nuclear exclusion of the Mcm2-7 complex. (A) Net nuclear export of NLS2-NLS3NES3-GFP₃ is dependent on the consensus CDK phosphorylation sites in NLS3NES3. YJL4662 (crm1Δ trp1::{pGAL-NLS2-NLS3NES3-GFP₃),TRP1} [crm1-T539C]), YJL5750 (trp1::{pGAL-NLS2-NLS3NES3-cdk5A-GFP₃),TRP1}) and YJL5753 (trp1::{pGAL-NLS2-NLS3NES3-cdk4A-GFP₃), TRP1}) growing exponentially and constitutively expressing their GFP₃ fusion proteins in YEPGal medium were examined by fluorescence microscopy. (B) Mcm3 CDK consensus sites are required for nuclear exclusion of Mcm3. Fluorescent microscopy of nocodazole-arrested YJL2714 (GFP-mcm3-cdk4A), YJL2720 (GFP-mcm3-cdk5A), and YJL2314(GFP-mcm3-cdk7A). (C) Mcm3 Cdk consensus sites are required for nuclear exclusion of other Mcm subunits. Fluorescence microscopy of nocodazole-arrested YJL2033 (mcm2::{MCM2-GFP}), YJL4165 (mcm2::{MCM2-GFP} mcm3-cdk5A), YJL2160 (GFP-MCM3), YJL2720 (GFP-mcm3-cdk5A), YJL2037 (mcm4::{MCM4-GFP}), YJL4169 (mcm4::{MCM4-GFP}, mcm3-cdk5A), YJL2217 (mcm7::{MCM7-GFP}), and YJL4167 (mcm7:: {MCM7-GFP}, mcm3-cdk5A).