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Related Concept Videos

DNA Damage can Stall the Cell Cycle02:36

DNA Damage can Stall the Cell Cycle

In response to DNA damage, cells can pause the cell cycle to assess and repair the breaks. However, the cell must check the DNA at certain critical stages during the cell cycle. If the cell cycle pauses before DNA replication, the cells will contain twice the amount of DNA. On the other hand, if cells arrest after DNA replication but before mitosis, they will contain four times the normal amount of DNA. With a host of specialized proteins at their disposal,cells must use the right protein at...
DNA Damage Can Stall the Cell Cycle02:36

DNA Damage Can Stall the Cell Cycle

In response to DNA damage, cells can pause the cell cycle to assess and repair the breaks. However, the cell must check the DNA at certain critical stages during the cell cycle. If the cell cycle pauses before DNA replication, the cells will contain twice the amount of DNA. On the other hand, if cells arrest after DNA replication but before mitosis, they will contain four times the normal amount of DNA. With a host of specialized proteins at their disposal,cells must use the right protein at...
Allosteric Proteins-ATCase01:19

Allosteric Proteins-ATCase

Binding sites linkages can regulate a protein's function.  For example, enzyme activity is often regulated through a feedback mechanism where the end product of the biochemical process serves as an inhibitor.
Aspartate transcarbamoylase (ATCase) is a cytosolic enzyme that catalyzes the condensation of L-aspartate and carbamoyl phosphate to  N-carbamoyl-L-aspartate. This reaction is the first step in pyrimidine biosynthesis. UTP and CTP, the end products of the pyrimidine synthesis pathway,...
The Spindle Assembly Checkpoint02:19

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...
The Unfolded Protein Response01:37

The Unfolded Protein Response

The ER is the hub of protein synthesis in a cell. It has robust systems to quality control protein folding and also for degradation of terminally misfolded proteins. Under normal conditions, a small proportion of misfolded proteins that cannot be salvaged need to be transported to the cytoplasm by the ER-associated degradation or ERAD pathways. However, if the ERAD cannot handle the misfolded proteins, the cell activates the unfolded protein response or UPR to adjust the protein folding...
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The Intrinsic Apoptotic Pathway

Internal cellular stress, such as cellular injury or hypoxia, triggers intrinsic apoptosis. The B-cell lymphoma 2 (Bcl-2) family of proteins are the primary regulators of the intrinsic apoptotic pathway. For example, during DNA damage, checkpoint proteins, such as Ataxia Telangiectasia Mutated (ATM protein) and Checkpoints Factor-2 (Chk2) proteins, are activated. These proteins phosphorylate p53 which further activates pro-apoptotic proteins, such as Bax, Bak, PUMA, and Noxa, and inhibits...

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Related Experiment Video

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Study of the DNA Damage Checkpoint using Xenopus Egg Extracts
10:55

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Published on: November 5, 2012

Non-catalytic function for ATR in the checkpoint response.

Troy D McSherry1, Ana A Kitazono, Ali Javaheri

  • 1Center for Molecular Oncology, University of Chicago, Chicago, Illinois, USA.

Cell Cycle (Georgetown, Tex.)
|August 28, 2007
PubMed
Summary

The non-catalytic portion of ATR kinases is crucial for DNA damage response. This conserved function involves the spindle assembly checkpoint, linking ATR to Mad1 and Mad2, independent of its catalytic activity.

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Area of Science:

  • Cell Biology
  • Genetics
  • Molecular Biology

Background:

  • The ATR (Ataxia Telangiectasia and Rad3-related) kinase family is vital for eukaryotic DNA damage response.
  • ATR kinases orchestrate cell cycle arrest, DNA repair, and apoptosis following genomic insults.
  • The catalytic domain is a small part of these large kinases, suggesting non-catalytic regions have functions.

Purpose of the Study:

  • To investigate the conserved function of the non-catalytic portion of ATR kinases.
  • To determine the role of Xenopus ATR (XATR) in Saccharomyces cerevisiae mec1 mutants.
  • To elucidate the specific checkpoint pathways involved in ATR's function.

Main Methods:

  • Expression of wild-type and kinase-defective XATR in S. cerevisiae mec1 mutants.
  • Analysis of checkpoint defects, cell cycle arrest, and dependence on specific yeast factors (Ddc2, Rad9, Rad53, Mad1, Mad2).
  • Creation and analysis of a yeast strain with a truncated mec1 lacking the kinase domain.

Main Results:

  • Xenopus ATR (XATR) suppressed the mec1 mutant checkpoint defect, inducing DNA damage-dependent mitotic arrest.
  • Suppression required yeast Ddc2 and Rad9 but was independent of Rad9 modification and Rad53 activation.
  • XATR-mediated suppression involved spindle checkpoint factors Mad1 and Mad2, suggesting a role in the spindle assembly checkpoint.
  • A truncated mec1 mutant was partially checkpoint proficient and induced Mad2-dependent mitotic arrest.

Conclusions:

  • The non-catalytic portion of ATR kinases has a conserved function in checkpoint control.
  • ATR kinases are linked to the spindle assembly checkpoint pathway via their non-catalytic regions.
  • This connection between ATR and the spindle checkpoint is conserved across species.