1. The authors studied this topic to investigate if the cohesion complex has any influence on chromosome instability and if it does, how to correct it. This is important because chromosome instability is proposed to be a major contributor to the acceleration of cancer cell evolution1, and as a previously overlooked source, cohesion fatigue can induce chromosome instability. The authors focused on the cohesion complex in this study since in a previous study they identified that a certain protein, Ska3, was important in the maintaining of chromosome cohesion2. This implies that there is a possibility of other proteins that can destabilize chromosome cohesion, thus creating chromosome instability.
2. Cohesion fatigue is a phenomenon named by the authors, in which during induced metaphase delay, spindle pulling forces cause asynchronous chromatid separation3. Cohesion fatigue was determined experimentally by arresting a cell’s metaphase through the use of proteasome inhibitors (MG132) and visualizing the chromatids separation through video microscopy.
3. SKA3 (previously named C13orf3) is a protein which plays a large role in the attachment of kinetochore-microtubules and movement of chromosomes2. SKA3 localizes to the spindle and kinetochore–microtubule interface throughout mitosis, …show more content…
Beta actin is a cytoskeletal protein involved in cell structure, motility, and cohesion. Also, it is highly conserved, which allows it to be constitutively expressed in all proteins, making it a good loading control6. This protein was used by the authors to indicate that when the separase RNAi and SKA3 RNAi were added to the cell, the western blot indicated this by having little to no bands for the amount of separase and SKA3 in the depleted cell. Since actin is constitutively expressed, as long as its band is present, it is clear if the other selected proteins are expressed or depleted in the cell, if they are loaded correctly and if the proteins are being transferred
2. Cohesion fatigue is a phenomenon named by the authors, in which during induced metaphase delay, spindle pulling forces cause asynchronous chromatid separation3. Cohesion fatigue was determined experimentally by arresting a cell’s metaphase through the use of proteasome inhibitors (MG132) and visualizing the chromatids separation through video microscopy.
3. SKA3 (previously named C13orf3) is a protein which plays a large role in the attachment of kinetochore-microtubules and movement of chromosomes2. SKA3 localizes to the spindle and kinetochore–microtubule interface throughout mitosis, …show more content…
Beta actin is a cytoskeletal protein involved in cell structure, motility, and cohesion. Also, it is highly conserved, which allows it to be constitutively expressed in all proteins, making it a good loading control6. This protein was used by the authors to indicate that when the separase RNAi and SKA3 RNAi were added to the cell, the western blot indicated this by having little to no bands for the amount of separase and SKA3 in the depleted cell. Since actin is constitutively expressed, as long as its band is present, it is clear if the other selected proteins are expressed or depleted in the cell, if they are loaded correctly and if the proteins are being transferred