Tetraploidy, a common feature on cancer, results in the presence of extra centrosomes, which have been associated with chromosomal instability (CIN) and aneuploidy. Deregulation of the number of centrosomes trigger tumorigenesis. However, what supernumerary centrosomes evolved during the rise of tetraploid cells is still not explained.
Here, generate isogenic tetraploid clones in colorectal cancer and non-transformed cells, we showed that the tetraploid near-clones showed a significant increase in the number of centrosomes. In addition, we found that the centrosome area near-tetraploids is twice as large in the near-diploid. To evaluate whether centrosome clustering that happens, we analyzed the number of centrioles reveal amplification centrioles.
Although more than half of the near-tetraploids maintained in culture centrosome aberrations not present. To test whether the cells are increasingly lost centrioles after being near-tetraploid, we transiently transfected diploid cells with siRNA against ESPL1 / Separase, protease responsible for triggering anaphase, to generate new cells in near-tetraploid. Finally, using this model, we assess the number of centrioles at different time-points after tetraploidization found that near-tetraploids quickly centrosomes lose from time to time.
Taken together, these data indicate that although the majority of the cells reduces the supernumerary centrosomes after tetraploidization, a small percentage remained extra centrioles, potentially resulting in CIN.
Centrosome reduction in newly-generated tetraploid cancer cells obtained by separase depletion
Dun1, Chk2 kinase-related, is the central regulator of securin-separase dynamics during DNA damage signal.
DNA damage checkpoint stop cycle progression at G2 cells in response to genotoxic insults. Central to the implementation of cell cycle checkpoint arrest is induced stabilization securin-separase complex (yeast Pds1-Esp1). checkpoint kinase Chk1 and Chk2 (yeast Chk1 and Rad53) is expected to contribute to the stability of the critical-separase securin complex by phosphorylation securin, rendering it resistant to proteolytic breakdown by the anaphase promoting complex (APC).
Dun1, a Rad53 paralog associated with Chk2, are also important for the post-imposed arrest. Dun1 required for transcription-induced DNA damage DNA repair genes; However, its role in the implementation of cell cycle arrest is still unknown. Here, we show that Dun1 role in the arrest checkpoint independent of its involvement in the improvement of gene transcription.
Instead, Dun1 Pds1 necessary to prevent damage during DNA damage in cells Dun1-deficiency lowers Pds1, escape G2 arrest and undergoing mitotic checkpoint despite the presence of active Chk1 and Rad53. Interestingly, the proteolytic degradation of Pds1 the absence Dun1 mediated not by APC but by HECT domain containing E3 ligase Rsp5. Our results demonstrate regulatory scheme where Dun1 prevent chromosome segregation during DNA damage by inhibiting Rsp5-mediated proteolytic degradation of securin Pds1.
Description: A polyclonal antibody for detection of Separase from Human, Mouse. This Separase antibody is for WB, IHC-P, IF, ELISA. It is affinity-purified from rabbit antiserum by affinity-chromatography using epitope-specific immunogenand is unconjugated. The antibody is produced in rabbit by using as an immunogen synthesized peptide derived from human Separase around the non-phosphorylation site of S801
Description: A polyclonal antibody for detection of Separase from Human, Mouse. This Separase antibody is for WB, IHC-P, IF, ELISA. It is affinity-purified from rabbit antiserum by affinity-chromatography using epitope-specific immunogenand is unconjugated. The antibody is produced in rabbit by using as an immunogen synthesized peptide derived from human Separase around the non-phosphorylation site of S801
Description: A polyclonal antibody for detection of Separase from Human, Mouse. This Separase antibody is for WB, IHC-P, IF, ELISA. It is affinity-purified from rabbit antiserum by affinity-chromatography using epitope-specific immunogenand is unconjugated. The antibody is produced in rabbit by using as an immunogen synthesized peptide derived from human Separase around the non-phosphorylation site of S801
Description: Stable cohesion between sister chromatids before anaphase and their timely separation during anaphase are critical for chromosome inheritance. In vertebrates, sister chromatid cohesion is released in 2 steps via distinct mechanisms. The first step involves phosphorylation of STAG1 (MIM 604358) or STAG2 (MIM 300826) in the cohesin complex. The second step involves cleavage of the cohesin subunit SCC1 (RAD21; MIM 606462) by ESPL1, or separase, which initiates the final separation of sister chromatids.
Description: Stable cohesion between sister chromatids before anaphase and their timely separation during anaphase are critical for chromosome inheritance. In vertebrates, sister chromatid cohesion is released in 2 steps via distinct mechanisms. The first step involves phosphorylation of STAG1 (MIM 604358) or STAG2 (MIM 300826) in the cohesin complex. The second step involves cleavage of the cohesin subunit SCC1 (RAD21; MIM 606462) by ESPL1, or separase, which initiates the final separation of sister chromatids.
Description: Stable cohesion between sister chromatids before anaphase and their timely separation during anaphase are critical for chromosome inheritance. In vertebrates, sister chromatid cohesion is released in 2 steps via distinct mechanisms. The first step involves phosphorylation of STAG1 (MIM 604358) or STAG2 (MIM 300826) in the cohesin complex. The second step involves cleavage of the cohesin subunit SCC1 (RAD21; MIM 606462) by ESPL1, or separase, which initiates the final separation of sister chromatids.
Description: A polyclonal antibody raised in Rabbit that recognizes and binds to Human ESPL1 / Separase (aa767-816). This antibody is tested and proven to work in the following applications:
Description: A polyclonal antibody for detection of Separase phospho Ser801) from Human, Mouse. This Separase phospho Ser801) antibody is for WB , IHC-P, IF, ELISA. It is affinity-purified from rabbit antiserum by affinity-chromatography using epitope-specific immunogenand is unconjugated. The antibody is produced in rabbit by using as an immunogen synthesized peptide derived from human Separase around the phosphorylation site of S801
Description: A polyclonal antibody for detection of Separase phospho Ser801) from Human, Mouse. This Separase phospho Ser801) antibody is for WB , IHC-P, IF, ELISA. It is affinity-purified from rabbit antiserum by affinity-chromatography using epitope-specific immunogenand is unconjugated. The antibody is produced in rabbit by using as an immunogen synthesized peptide derived from human Separase around the phosphorylation site of S801
Description: A polyclonal antibody for detection of Separase phospho Ser801) from Human, Mouse. This Separase phospho Ser801) antibody is for WB , IHC-P, IF, ELISA. It is affinity-purified from rabbit antiserum by affinity-chromatography using epitope-specific immunogenand is unconjugated. The antibody is produced in rabbit by using as an immunogen synthesized peptide derived from human Separase around the phosphorylation site of S801
Description: Stable cohesion between sister chromatids before anaphase and their timely separation during anaphase are critical for chromosome inheritance. In vertebrates| sister chromatid cohesion is released in 2 steps via distinct mechanisms. The first step involves phosphorylation of STAG1 (MIM 604358) or STAG2 (MIM 300826) in the cohesin complex. The second step involves cleavage of the cohesin subunit SCC1 (RAD21; MIM 606462) by ESPL1| or separase| which initiates the final separation of sister chromatids (Sun et al.| 2009
Description: Stable cohesion between sister chromatids before anaphase and their timely separation during anaphase are critical for chromosome inheritance. In vertebrates| sister chromatid cohesion is released in 2 steps via distinct mechanisms. The first step involves phosphorylation of STAG1 (MIM 604358) or STAG2 (MIM 300826) in the cohesin complex. The second step involves cleavage of the cohesin subunit SCC1 (RAD21; MIM 606462) by ESPL1| or separase| which initiates the final separation of sister chromatids (Sun et al.| 2009
Description: Stable cohesion between sister chromatids before anaphase and their timely separation during anaphase are critical for chromosome inheritance. In vertebrates| sister chromatid cohesion is released in 2 steps via distinct mechanisms. The first step involves phosphorylation of STAG1 (MIM 604358) or STAG2 (MIM 300826) in the cohesin complex. The second step involves cleavage of the cohesin subunit SCC1 (RAD21; MIM 606462) by ESPL1| or separase| which initiates the final separation of sister chromatids (Sun et al.| 2009
Description: A Rabbit Polyclonal antibody against Separase (phospho Ser801) from Human/Mouse. This antibody is tested and validated for WB, ELISA, IHC, IF, WB, ELISA
Description: A Rabbit Polyclonal antibody against Separase (phospho Ser801) from Human/Mouse. This antibody is tested and validated for WB, ELISA, IHC, IF, WB, ELISA
Often resulting in prolonged mitotic apoptosis1. Abbreviated tumorigenic mitosis causing aneuploidy, but it is not clear whether it also activates apoptosis machinery2. Separase, cysteine proteases and triggers of all anaphases eukaryotic, have caspase-like catalytic domains but have never been associated with cell death3,4. Here we show that human cells entering mitosis with separase already active to quickly undergo mitosis death because cleavage directly MCL1 and anti-apoptotic BCL-XL by separase. Cleavage is not only prevent MCL1 and BCL-XL from the execution of pro-apoptotic BAK, but also convert them into active promoters of death in mitosis.
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