The parafibromin tumor suppressor protein interacts with actin-binding proteins actinin-2 and actinin-3.
BACKGROUND: Germline and somatic inactivating mutations in the HRPT2 gene occur in the inherited hyperparathyroidism-jaw tumor syndrome, in some cases of parathyroid cancer and in some cases of familial hyperparathyroidism. HRPT2 encodes parafibromin. To identify parafibromin interacting proteins we used the yeast two-hybrid system for screening a heart cDNA library ... with parafibromin as the bait. RESULTS: Fourteen parafibromin interaction positive preys representing 10 independent clones encoding actinin-2 were isolated. Parafibromin interacted with muscle alpha-actinins (actinin-2 and actinin-3), but not with non-muscle alpha-actinins (actinin-1 and actinin-4). The parafibromin-actinin interaction was verified by yeast two-hybrid, GST pull-down, and co-immunoprecipitation. Yeast two-hybrid analysis revealed that the N-terminal region of parafibromin interacted with actinins. In actin sedimentation assays parafibromin did not dissociate skeletal muscle actinins from actin filaments, but interestingly, parafibromin could also bundle/cross-link actin filaments. Parafibromin was predominantly nuclear in undifferentiated proliferating myoblasts (C2C12 cells), but in differentiated C2C12 myotubes parafibromin co-localized with actinins in the cytoplasmic compartment. CONCLUSION: These data support a possible contribution of parafibromin outside the nucleus through its interaction with actinins and actin bundling/cross-linking. These data also suggest that actinins (and actin) participate in sequestering parafibromin in the cytoplasmic compartment.
Mesh Terms:
Actinin, Animals, Cell Nucleus, Cells, Cultured, Cytoplasm, Fluorescent Antibody Technique, Gene Library, Hela Cells, Humans, Mice, Parathyroid Neoplasms, Transfection, Tumor Suppressor Proteins, Two-Hybrid System Techniques
Actinin, Animals, Cell Nucleus, Cells, Cultured, Cytoplasm, Fluorescent Antibody Technique, Gene Library, Hela Cells, Humans, Mice, Parathyroid Neoplasms, Transfection, Tumor Suppressor Proteins, Two-Hybrid System Techniques
Mol. Cancer
Date: Aug. 09, 2008
PubMed ID: 18687124
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