Related Experiment Video

Updated: Nov 22, 2025

Visualization of DNA Repair Proteins Interaction by Immunofluorescence
07:55

Visualization of DNA Repair Proteins Interaction by Immunofluorescence

Published on: June 26, 2020

10.7K

A Spatial and Functional Interaction of a Heterotetramer Survivin-DNA-PKcs Complex in DNA Damage Response

Ömer Güllülü1, Stephanie Hehlgans1, Benjamin E Mayer2

  • 1Department of Radiotherapy and Oncology, University Hospital, Goethe University Frankfurt, Germany.

Cancer Research
|January 7, 2021
PubMed

Insights

Survivin protein overexpression enhances tumor radiation resistance by directly interacting with DNA-PKcs, altering DNA repair and promoting cancer survival. This study reveals the molecular mechanism behind survivin's role in DNA damage response.

Area of Science:

  • Molecular oncology
  • DNA damage response
  • Cancer biology

Background:

  • Overexpression of inhibitor of apoptosis protein (IAP) survivin in tumors correlates with treatment resistance and poor prognosis.
  • Survivin acts as a radiation resistance factor impacting DNA damage response via interaction with DNA-dependent protein kinase (DNA-PKcs).
  • The precise molecular mechanisms and functional outcomes of the survivin-DNA-PKcs relationship are not fully understood.

Purpose of the Study:

  • To elucidate the molecular determinants and functional consequences of survivin's interaction with DNA-PKcs in regulating radiation survival and DNA repair.
  • To investigate the structural basis and enzymatic activity changes in the DNA-PKcs-survivin complex.

Main Methods:

  • Coimmunoprecipitation and Förster resonance energy transfer (FRET) assays to demonstrate direct interaction.
  • In silico molecular docking and dynamics simulations.
  • In vitro kinase assays and large-scale mass spectrometry.
  • Overexpression studies in cancer cells.

Main Results:

  • Survivin directly interacts with DNA-PKcs through its baculovirus IAP repeat domain, specifically involving residues S20 and W67 with the PI3K domain of DNA-PKcs.
  • A heterotetrameric survivin-DNA-PKcs complex forms, inducing a conformational change in the DNA-PKcs PI3K domain and enhancing its enzymatic activity.
  • Survivin alters DNA-PKcs substrate specificity, favoring S/T-P phosphorylation sites and increasing substrates like Foxo3, thereby modulating DNA double-strand break repair and radiation survival.

Conclusions:

  • Survivin differentially regulates DNA-PKcs-dependent radiation survival and DNA double-strand break repair through the formation of a heterotetrameric complex.
  • These findings provide critical insights into survivin's role in modulating the cellular radiation response and DNA repair pathways.
  • The study highlights survivin as a key regulator of DNA-PKcs kinase activity in the context of cancer treatment resistance.

More Related Videos

Detection and Visualization of DNA Damage-induced Protein Complexes in Suspension Cell Cultures Using the Proximity Ligation Assay13:10

Detection and Visualization of DNA Damage-induced Protein Complexes in Suspension Cell Cultures Using the Proximity Ligation Assay

Published on: June 9, 2017

10.3K
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

14.4K

Related Concept Videos

DNA Damage can Stall the Cell Cycle02:37

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...
9.7K
DNA Damage Can Stall the Cell Cycle02:37

DNA Damage Can Stall the Cell Cycle

2.9K
Homologous Recombination02:31

Homologous Recombination

The basic reaction of homologous recombination (HR) involves two chromatids that contain DNA sequences sharing a significant stretch of identity. One of these sequences uses a strand from another as a template to synthesize DNA in an enzyme-catalyzed reaction. The final product is a novel amalgamation of the two substrates. To ensure an accurate recombination of sequences, HR is restricted to the S and G2 phases of the cell cycle. At these stages, the DNA has been replicated already and the...
59.4K
Homologous Recombination02:31

Homologous Recombination

5.5K
Fixing Double-strand Breaks02:04

Fixing Double-strand Breaks

The double-stranded structure of DNA has two major advantages. First, it serves as a safe repository of genetic information where one strand serves as the back-up in case the other strand is damaged. Second, the double-helical structure can be wrapped around proteins called histones to form nucleosomes, which can then be tightly wound to form chromosomes. This way, DNA chains up to 2 inches long can be contained within microscopic structures in a cell. A double-stranded break not only damages...
13.5K
Restarting Stalled Replication Forks02:37

Restarting Stalled Replication Forks

DNA replication is initiated at sites containing predefined DNA sequences known as origins of replication. DNA is unwound at these sites by the minichromosome maintenance (MCM) helicase and other factors such as Cdc45 and the associated GINS complex.The unwound single strands are protected by replication protein A (RPA) until DNA polymerase starts synthesizing DNA at the 5’ end of the strand in the same direction as the replication fork. To prevent the replication fork from falling apart,...
6.1K