Functional Interactions between BM88/Cend1, Ran-Binding Protein M and Dyrk1B Kinase Affect Cyclin D1 Levels and Cell Cycle Progression/Exit in Mouse Neuroblastoma Cells
Figure 1
Schematic representation of Cend1 and RanBPM cDNA constructs and protein fragments.
(a) Total mouse Cend1 cDNA (1241-bp) encodes for a 149 amino-acid protein, containing a C-terminal transmembrane region (TM) followed by RKK tail. A 375 bp fragment (372 bp plus a stop codon) corresponding to nucleotides 141 to 513 of Cend1 cDNA) and encoding the first 124 amino acids of the protein, was generated by high-fidelity PCR. This fragment was fused in frame with either the GAL4 binding domain or GST and served for yeast two hybrid screening or GST-pull down assays, respectively. (b, c) Diagrammatic illustration of the three RanBPM cDNA clones (b) and corresponding protein fragments (c) isolated from yeast two hybrid screening, all containing the multifunctional proline-rich SPRY-LISH-CTLH-CRA domain. Generation of GST-RanBPM chimeric molecules (corresponding to the 981 bp and 1731 bp cDNA fragments, as indicated) and the SPRY-LISH-CTLH domain (942 bp) was performed by high-fidelity PCR.