1. AKAP95 interacts with nucleoporin TPR in mitosis and is important for the spindle assembly checkpoint.
- Author
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López-Soop G, Rønningen T, Rogala A, Richartz N, Blomhoff HK, Thiede B, Collas P, and Küntziger T
- Subjects
- Chromosome Segregation genetics, HeLa Cells, Humans, Mitosis genetics, Nuclear Pore genetics, Nuclear Pore metabolism, Nuclear Pore Complex Proteins metabolism, Protein Binding, Spindle Apparatus genetics, Spindle Apparatus metabolism, A Kinase Anchor Proteins genetics, M Phase Cell Cycle Checkpoints genetics, Nuclear Pore Complex Proteins genetics, Proteomics, Proto-Oncogene Proteins genetics
- Abstract
Faithful chromosome segregation during mitosis relies on a proofreading mechanism that monitors proper kinetochore-microtubule attachments. The spindle assembly checkpoint (SAC) is based on the concerted action of numerous components that maintain a repressive signal inhibiting transition into anaphase until all chromosomes are attached. Here we show that A-Kinase Anchoring Protein 95 (AKAP95) is necessary for proper SAC function. AKAP95-depleted HeLa cells show micronuclei formed from lagging chromosomes at mitosis. Using a BioID proximity-based proteomic screen, we identify the nuclear pore complex protein TPR as a novel AKAP95 binding partner. We show interaction between AKAP95 and TPR in mitosis, and an AKAP95-dependent enrichment of TPR in the spindle microtubule area in metaphase, then later in the spindle midzone area. AKAP95-depleted cells display faster prometaphase to anaphase transition, escape from nocodazole-induced mitotic arrest and show a partial delocalization from kinetochores of the SAC component MAD1. Our results demonstrate an involvement of AKAP95 in proper SAC function likely through its interaction with TPR.
- Published
- 2017
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