Related Experiment Video

Updated: Apr 6, 2026

Analysis of Cell Cycle Position in Mammalian Cells
12:19

Analysis of Cell Cycle Position in Mammalian Cells

Published on: January 21, 2012

61.6K

Assessing Cell Cycle Independent Function of the CDK Inhibitor p21(CDKN¹A) in DNA Repair

Ilaria Dutto1, Micol Tillhon, Ennio Prosperi

  • 1Genome Stability Group, Department of Biology and Biotechnology, Istituto di Genetica Molecolare del CNR, University of Pavia, Via Ferrata 1/9, 27100, Pavia, Italy.

Methods in Molecular Biology (Clifton, N.J.)
|August 2, 2015
PubMed

Insights

The cyclin-dependent kinase (CDK) inhibitor p21 (CDKN1A) plays a crucial role in DNA repair, independently of its cell cycle inhibition. This study details methods to investigate p21

Area of Science:

  • Molecular Biology
  • Cell Biology
  • Biochemistry

Background:

  • The cyclin-dependent kinase (CDK) inhibitor p21 (CDKN1A) regulates critical cellular functions beyond cell cycle control, including DNA repair.
  • p21's involvement in DNA repair pathways like NER, BER, and DSB repair is established through its interaction with proteins such as PCNA.
  • The precise mechanism linking p21's DNA repair role to its proteasomal degradation post-DNA damage requires further elucidation.

Purpose of the Study:

  • To establish and describe experimental procedures for investigating the recruitment of p21 protein to localized DNA damage sites within the cell nucleus.
  • To provide a comprehensive guide for studying p21's function in DNA repair and its interactions with other cellular components.

Main Methods:

  • Localized DNA damage induction using UV irradiation through a microporous polycarbonate filter.
  • In situ lysis followed by immunofluorescence to detect chromatin-bound proteins.
  • Cell fractionation and Western blot analysis to assess p21's chromatin association.
  • Immunoprecipitation assays to identify p21 protein-protein interactions.

Main Results:

  • The described methods enable the study of p21 recruitment to specific DNA damage sites.
  • These techniques allow for the analysis of p21's association with chromatin and its interactions with other proteins involved in DNA repair.
  • The procedures facilitate the investigation of how p21 functions in DNA repair pathways.

Conclusions:

  • Established protocols allow for the detailed study of p21 protein dynamics at DNA damage sites.
  • These methods are essential for understanding the established role of p21 in DNA repair mechanisms.
  • Further research using these techniques can clarify the interplay between p21's DNA repair functions and its degradation pathways.

More Related Videos

Two- and Three-Dimensional Live Cell Imaging of DNA Damage Response Proteins10:24

Two- and Three-Dimensional Live Cell Imaging of DNA Damage Response Proteins

Published on: September 28, 2012

14.7K
Assessment of Global DNA Double-Strand End Resection using BrdU-DNA Labeling coupled with Cell Cycle Discrimination Imaging06:44

Assessment of Global DNA Double-Strand End Resection using BrdU-DNA Labeling coupled with Cell Cycle Discrimination Imaging

Published on: April 28, 2021

4.7K

Related Concept Videos

Inhibition of Cdk Activity02:34

Inhibition of Cdk Activity

The orderly progression of the cell cycle depends on the activation of Cdk protein by binding to its cyclin partner. However, the cell cycle must be restricted when undergoing abnormal changes. Most cancers correlate to the deregulated cell cycle, and since Cdks are a central component of the cell cycle, Cdk inhibitors are extensively studied to develop anticancer agents. For instance, cyclin D associates with several Cdks, such as Cdk 4/6, to form an active complex. The cyclin D-Cdk4/6 complex...
6.2K
Inhibition of CDK Activity02:34

Inhibition of CDK Activity

5.7K
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...
3.4K
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...
10.4K
Negative Regulator Molecules01:23

Negative Regulator Molecules

Positive regulators allow a cell to advance through cell cycle checkpoints. Negative regulators have an equally important role as they terminate a cell’s progression through the cell cycle—or pause it—until the cell meets specific criteria.
39.0K
Positive Regulator Molecules02:39

Positive Regulator Molecules

Mitotic cell division results in daughter cells that exactly resemble the parent cell. However, errors in the DNA replication or distribution of genetic material may lead to genetic mutations that may be passed down to every new cell formed from the resulting abnormal cell. Propagation of such mutant cells is restricted through checkpoint mechanisms present at different stages of the cell cycle. These checkpoints involve regulator molecules that either promote or demote cell cycle events.
7.1K