PMID- 10572166
OWN - NLM
STAT- MEDLINE
DCOM- 20000125
LR  - 20190501
IS  - 1362-4962 (Electronic)
IS  - 0305-1048 (Linking)
VI  - 27
IP  - 24
DP  - 1999 Dec 15
TI  - Geometry of a complex formed by double strand break repair proteins at a single
      DNA end: recruitment of DNA-PKcs induces inward translocation of Ku protein.
PG  - 4679-86
AB  - Ku protein and the DNA-dependent protein kinase catalytic subunit (DNA-PKcs) are 
      essential components of the double-strand break repair machinery in higher
      eukaryotic cells. Ku protein binds to broken DNA ends and recruits DNA-PKcs to
      form an enzymatically active complex. To characterize the arrangement of proteins
      in this complex, we developed a set of photocross-linking probes, each with a
      single free end. We have previously used this approach to characterize the
      contacts in an initial Ku-DNA complex, and we have now applied the same
      technology to define the events that occur when Ku recruits DNA-PKcs. The new
      probes allow the binding of one molecule of Ku protein and one molecule of
      DNA-PKcs in a defined position and orientation. Photocross-linking reveals that
      DNA-PKcs makes direct contact with the DNA termini, occupying an approximately 10
      bp region proximal to the free end. Characterization of the Ku protein
      cross-linking pattern in the presence and absence of DNA-PKcs suggests that Ku
      binds to form an initial complex at the DNA ends, and that recruitment of
      DNA-PKcs induces an inward translocation of this Ku molecule by about one helical
      turn. The presence of ATP had no effect on protein-DNA contacts, suggesting that 
      neither DNA-PK-mediated phosphorylation nor a putative Ku helicase activity plays
      a role in modulating protein conformation under the conditions tested.
FAU - Yoo, S
AU  - Yoo S
AD  - Program in Gene Regulation, Institute of Molecular Medicine and Genetics, Medical
      College of Georgia, 1120 15th Street, Augusta, GA 30912, USA.
FAU - Dynan, W S
AU  - Dynan WS
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PL  - England
TA  - Nucleic Acids Res
JT  - Nucleic acids research
JID - 0411011
RN  - 0 (Antigens, Nuclear)
RN  - 0 (Cross-Linking Reagents)
RN  - 0 (DNA Probes)
RN  - 0 (DNA-Binding Proteins)
RN  - 0 (Nuclear Proteins)
RN  - 9007-49-2 (DNA)
RN  - EC 2.7.11.1 (DNA-Activated Protein Kinase)
RN  - EC 2.7.11.1 (PRKDC protein, human)
RN  - EC 2.7.11.1 (Protein-Serine-Threonine Kinases)
RN  - EC 3.6.4.- (DNA Helicases)
RN  - EC 3.6.4.12 (XRCC5 protein, human)
RN  - EC 3.6.4.12 (Xrcc6 protein, human)
RN  - EC 4.2.99.- (Ku Autoantigen)
SB  - IM
MH  - *Antigens, Nuclear
MH  - Base Sequence
MH  - Binding Sites
MH  - Cell Nucleus/metabolism
MH  - Cross-Linking Reagents
MH  - DNA/*chemistry/*metabolism
MH  - DNA Damage
MH  - *DNA Helicases
MH  - DNA Probes/chemistry
MH  - *DNA Repair
MH  - DNA-Activated Protein Kinase
MH  - DNA-Binding Proteins/*chemistry/*metabolism
MH  - HeLa Cells
MH  - Humans
MH  - Kinetics
MH  - Ku Autoantigen
MH  - Models, Molecular
MH  - Nuclear Proteins/*chemistry/*metabolism
MH  - Nucleic Acid Conformation
MH  - Protein Structure, Quaternary
MH  - Protein-Serine-Threonine Kinases/*chemistry/*metabolism
PMC - PMC148766
EDAT- 1999/11/26 00:00
MHDA- 1999/11/26 00:01
CRDT- 1999/11/26 00:00
PHST- 1999/11/26 00:00 [pubmed]
PHST- 1999/11/26 00:01 [medline]
PHST- 1999/11/26 00:00 [entrez]
AID - gkc692 [pii]
AID - 10.1093/nar/27.24.4679 [doi]
PST - ppublish
SO  - Nucleic Acids Res. 1999 Dec 15;27(24):4679-86. doi: 10.1093/nar/27.24.4679.