相关实验视频

Updated: Jul 1, 2025

Immunofluorescence Analysis of Endogenous and Exogenous Centromere-kinetochore Proteins

Immunofluorescence Analysis of Endogenous and Exogenous Centromere-kinetochore Proteins

Published on: March 3, 2016

通过调节微管-染色体连接,CKAP5稳定了CENP-E在基因中

R Bhagya Lakshmi¹, Pinaki Nayak², Linoy Raz³

¹School of Biology, Indian Institute of Science Education and Research, Thiruvananthapuram, Vithura, Thiruvananthapuram, Kerala, 695551, India.

|February 29, 2024

概括

CKAP5蛋白对于正确的染色体对齐至关重要,它通过稳定CENP-E在kinetochores. 它的耗尽会导致错误,特别是在染色体不稳定的癌细胞中.

科学领域:

细胞生物学细胞生物学
分子生物学分子生物学
遗传学是一个遗传学.

背景情况:

在细胞分裂过程中,基因 - 微管附着物对于精确的染色体分离至关重要.
基因素运动蛋白CENP-E在稳定基因基因基因基因基因基因基因基因基因基因基因基因基因基因基因基因基因基因基因基因基因基因基因基因基因基因基因.
准确的机制调节CENP-E稳定在kinetochores仍然在很大程度上是未知的.

研究的目的:

为了研究CKAP5的作用,一个微管加尖端蛋白,在调节CENP-E功能在kinetochores.
为了确定CKAP5如何影响染色体对齐和kinetochore-microtubule稳定性.
探索CKAP5在具有染色体不稳定的癌细胞中的关键性.

主要方法:

在人类细胞和癌症细胞系中使用遗传方法来消耗CKAP5.
免疫光显微镜以评估CENP-E局部化和动脉-微管附着物.
计算建模来模拟动态-微管子动态.
分析蛋白质与蛋白质相互作用 (CENP-E和BubR1) 和酸化.
对CKAP5淘汰癌细胞中的细胞活力和本质性的评估.

主要成果:

CKAP5的枯竭破坏了CENP-E在基因相基因的局部化,导致基因-微管稳定性增加和附着错误.
CKAP5在体,染色体不稳定的癌细胞中是首选必不可少的,其灵敏度与CENP-E枯竭相关.
CKAP5调节了CENP-E和BubR1之间的相互作用,其TOG4-TOG5域对这种相互作用和CENP-E稳定至关重要.
在CKAP5枯竭的细胞中,CKAP5枯竭促使PP1被引入基因,并且在CKAP5枯竭的细胞中稳定了一种模仿素的CENP-E突变体.

结论:

CKAP5 作为基因细胞中CENP-E的关键调节者,控制了线性染色体的附着忠实性.
CKAP5稳定了CENP-E,并调节了动态管附着微管的稳定性,从而防止了染色体分离错误.
在染色体不稳定的癌症中,CKAP5是潜在的治疗标,因为它在维护基因组完整性方面起着至关重要的作用.

关键词:

在CENP-E中,在CKAP5中,CKAP5是CKAP5.在Kinetochore中进行演出.微管是微管中的一个.PP1 PP1 PP1 PP1 PP1 PP1 PP1 PP1 PP1 PP1 PP1 PP1 PP1 PP1 PP1 PP1

更多相关视频

Author Spotlight: Establishing CENP-E Knockout HeLa Cells – A Novel Approach to Study Kinesin-7 CENP-E Biology and its Inhibitors

Author Spotlight: Establishing CENP-E Knockout HeLa Cells – A Novel Approach to Study Kinesin-7 CENP-E Biology and its Inhibitors

Published on: June 23, 2023

Identification of Cyclin-dependent Kinase 1 Specific Phosphorylation Sites by an In Vitro Kinase Assay

Identification of Cyclin-dependent Kinase 1 Specific Phosphorylation Sites by an In Vitro Kinase Assay

Published on: May 3, 2018

相关概念视频

Microtubule Instability

Microtubule Instability

Microtubules are hollow cylindrical filaments having a diameter of approximately 25 nm and a length that varies from 200 nm to 25 μm. GTP-bound tubulin subunits form αβ-heterodimers for microtubule assembly. These core building blocks interact longitudinally, polymerizing into protofilaments. The protofilaments then interact with one another through lateral bonding forces to form stable cylindrical microtubules. These cylindrical filaments are dynamic as they undergo repeated...

The Spindle Assembly Checkpoint

The Spindle Assembly Checkpoint

The spindle assembly checkpoint is a molecular surveillance mechanism ensuring the fidelity of chromosome segregation during anaphase. The checkpoint monitors the completion of all the prerequisite steps before chromosome segregation to determine whether the segregation process should proceed or be delayed.
Many proteins function together to control the spindle assembly checkpoint. Mutations affecting these proteins may allow cells to proceed into anaphase prematurely, resulting in the...

Microtubule Associated Proteins (MAPs)

Microtubule Associated Proteins (MAPs)

Microtubule function and architecture are regulated by an array of specialized proteins called microtubule-associated proteins or MAPs. These proteins are widespread across different organisms and have conserved protein motifs, like the multi-TOG domain for tubulin binding found in the CLASP family of MAPs. Some MAPs are lineage-specific based on their conserved domains. Their functions depend upon the cytoskeletal architecture and cell type they are located within. In-plant cells, a specific...

Separation of Sister Chromatids

Separation of Sister Chromatids

At the transition from prophase to metaphase, there is a reduction in cohesion along the chromosomal arms, resulting in the resolution of sister chromatids. However, residual cohesin connections remain to hold the sister chromatids together until the transition from metaphase to anaphase. The residual connection prevents any premature separation of sister chromatids, blocking the risks of aneuploidy within the daughter cells.
At the onset of anaphase, separase, a proteolytic enzyme, is...

Attachment of Sister Chromatids

Attachment of Sister Chromatids

As cells progress into mitosis, the nuclear envelope breaks down, and the condensed chromosomes are exposed to the array of bipolar microtubules of the mitotic spindle. The kinetochore, a large, disc-shaped protein complex, is present at the centromere region of the sister chromatids and acts as a binding site for the microtubules. Usually, the plus-end of a single microtubule is embedded within the kinetochore. However, some kinetochores first establish lateral contact with the side-wall...

Anaphase Promoting Complex

Anaphase Promoting Complex

The stepwise destruction of specific proteins is necessary for the progression and completion of the cell cycle. Such proteins are ubiquitinated by ubiquitin ligases and then subsequently destroyed by the proteasome. The SCF (Skp1/Cullin/F-box) and the anaphase-promoting complex (APC) are two important ubiquitin ligases involved in cell cycle progression. While SCF is active throughout the cell cycle, APC gets activated during metaphase to anaphase transition. Cdc20 or Cdh1 binds to APC and...