skip to main content
OSTI.GOV title logo U.S. Department of Energy
Office of Scientific and Technical Information

Title: Insights into Rad3 kinase recruitment from the crystal structure of the DNA damage checkpoint protein Rad26

Authors:
Publication Date:
Research Org.:
Argonne National Lab. (ANL), Argonne, IL (United States). Advanced Photon Source (APS)
Sponsoring Org.:
FOREIGN
OSTI Identifier:
1430298
Resource Type:
Journal Article
Resource Relation:
Journal Name: Journal of Biological Chemistry; Journal Volume: 292; Journal Issue: 20
Country of Publication:
United States
Language:
ENGLISH

Citation Formats

Andersen, Kasper Røjkjær. Insights into Rad3 kinase recruitment from the crystal structure of the DNA damage checkpoint protein Rad26. United States: N. p., 2017. Web. doi:10.1074/jbc.M117.780189.
Andersen, Kasper Røjkjær. Insights into Rad3 kinase recruitment from the crystal structure of the DNA damage checkpoint protein Rad26. United States. doi:10.1074/jbc.M117.780189.
Andersen, Kasper Røjkjær. Fri . "Insights into Rad3 kinase recruitment from the crystal structure of the DNA damage checkpoint protein Rad26". United States. doi:10.1074/jbc.M117.780189.
@article{osti_1430298,
title = {Insights into Rad3 kinase recruitment from the crystal structure of the DNA damage checkpoint protein Rad26},
author = {Andersen, Kasper Røjkjær},
abstractNote = {},
doi = {10.1074/jbc.M117.780189},
journal = {Journal of Biological Chemistry},
number = 20,
volume = 292,
place = {United States},
year = {Fri Mar 17 00:00:00 EDT 2017},
month = {Fri Mar 17 00:00:00 EDT 2017}
}
  • The CHK2 protein kinase is an important transducer of DNA damage checkpoint signals, and its mutation contributes to hereditary and sporadic cancer. CHK2 activation is triggered by the phosphorylation of Thr68 by the DNA damage-activated ATM kinase. This leads to transient CHK2 dimerization, in part through intermolecular phosphoThr68-FHA domain interactions. Dimerization promotes kinase activation through activation-loop autophosphorylation, but the mechanism of this process has not been clear. The dimeric crystal structure of CHK2, described here, in conjunction with biochemical and mutational data reveals that productive CHK2 dimerization additionally involves intermolecular FHA-kinase domain and FHA-FHA interactions. Ile157, mutated in the Li-Fraumenimore » cancer-predisposition syndrome, plays a central role in the FHA-kinase domain interface, explaining the lack of dimerization and autophosphorylation of this mutant. In the dimer, the kinase active sites face each other in close proximity, indicating that dimerization may also serve to optimally position the kinase active sites for efficient activation loop transphosphorylation.« less
  • In mitosis, the spindle checkpoint detects a single unattached kinetochore, inhibits the anaphase-promoting complex or cyclosome (APC/C), and prevents premature sister chromatid separation. The checkpoint kinase Bub1 contributes to checkpoint sensitivity through phosphorylating the APC/C activator, Cdc20, and inhibiting APC/C catalytically. We report here the crystal structure of the kinase domain of Bub1, revealing the requirement of an N-terminal extension for its kinase activity. Though the activation segment of Bub1 is ordered and has structural features indicative of active kinases, the C-terminal portion of this segment sterically restricts substrate access to the active site. Bub1 uses docking motifs, so-called KENmore » boxes, outside its kinase domain to recruit Cdc20, one of two known KEN box receptors. The KEN boxes of Bub1 are required for the spindle checkpoint in human cells. Therefore, its unusual active-site conformation and mode of substrate recruitment suggest that Bub1 has an exquisitely tuned specificity for Cdc20.« less
  • The DNA polymerase from phage {phi}29 is a B family polymerase that initiates replication using a protein as a primer, attaching the first nucleotide of the phage genome to the hydroxyl of a specific serine of the priming protein. The crystal structure of {phi}29 DNA polymerase determined at 2.2 Angstrom resolution provides explanations for its extraordinary processivity and strand displacement activities. Homology modeling suggests that downstream template DNA passes through a tunnel prior to entering the polymerase active site. This tunnel is too small to accommodate double-stranded DNA and requires the separation of template and nontemplate strands. Members of themore » B family of DNA polymerases that use protein primers contain two sequence insertions: one forms a domain not previously observed in polymerases, while the second resembles the specificity loop of T7 RNA polymerase. The high processivity of {phi}29 DNA polymerase may be explained by its topological encirclement of both the downstream template and the upstream duplex DNA.« less