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Published on: November 5, 2012
ATRIP oligomerization is required for ATR-dependent checkpoint signaling
1Department of Biochemistry, Vanderbilt University, Nashville, Tennessee 37232, USA.
The Journal of Biological Chemistry
|July 20, 2005
Summary
The ATR kinase and its ATRIP subunit form stable homooligomers essential for DNA damage response. ATRIP oligomerization, mediated by its coiled-coil domain, is crucial for ATR binding and checkpoint signaling.
Area of Science:
- Cellular biology
- Molecular genetics
- DNA damage response
Background:
- ATM and ATR kinases are crucial for cell cycle checkpoints following DNA damage.
- Inactive ATM functions as an oligomer, dissociating into active monomers upon DNA damage.
- The activation mechanism of ATR, particularly its oligomeric state, remains less understood.
Purpose of the Study:
- To investigate whether ATR activation involves a similar oligomer dissociation mechanism as ATM.
- To determine the oligomeric state of ATR and its subunit ATRIP.
- To elucidate the role of ATRIP oligomerization in ATR function and DNA damage signaling.
Main Methods:
- Immunoprecipitation and Western blotting to assess ATR and ATRIP oligomerization.
- Analysis of ATRIP mutants lacking the coiled-coil domain.
- Cellular localization studies using immunofluorescence microscopy.
- Assessment of checkpoint signaling to Chk1.
Main Results:
- ATR and ATRIP exist as stable homooligomers in cells.
- No significant regulation of ATR or ATRIP oligomerization was observed after DNA damage.
- The coiled-coil domain of ATRIP is critical for ATRIP oligomerization, stable ATR binding, and recruitment to DNA lesions.
- ATRIP oligomerization is necessary for ATR-dependent Chk1 signaling.
Conclusions:
- The ATR-ATRIP complex forms higher-order oligomeric structures.
- ATRIP oligomerization, independent of DNA damage, is essential for ATR complex stability and function in DNA damage response.
- The coiled-coil domain of ATRIP plays a pivotal role in mediating these functions.
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