DNA-dependent protein kinase (Ku protein-p350 complex) assembles on double-stranded DNA

Akira Suwa, Michito Hirakata, Yoshihiko Takeda, Stephen A. Jesch, Tsuneyo Mimori, John A. Hardin

Research output: Contribution to journalArticle

160 Citations (Scopus)

Abstract

The Ku protein is an autoantigen that consists of 70- and 80-kDa polypeptides. It associates with double-stranded DNA at free ends. In the present study, we examined the ability of anti-Ku antibodies to immunoprecipitate various structures from extracts of HeLa cells prepared at different salt concentrations. Under physiological conditions, these antibodies identified a complex containing the Ku protein and the 350-kDa component (p350) of DNA-dependent protein kinase (DNA-PK), which appeared to be closely associated on the DNA strand. In reconstitution experiments with cell extracts and biochemically purified components, the Ku protein-p350 complex formed only in the presence of double-stranded DNA. The reconstituted complex was catalytically active. Together with previous studies, these results indicate that the Ku protein interacts with DNA to create a binding site for p350 as the DNA-PK holoenzyme assembles.

Original languageEnglish (US)
Pages (from-to)6904-6908
Number of pages5
JournalProceedings of the National Academy of Sciences of the United States of America
Volume91
Issue number15
DOIs
StatePublished - Jul 19 1994
Externally publishedYes

Fingerprint

DNA-Activated Protein Kinase
DNA
Holoenzymes
Autoantigens
Cell Extracts
HeLa Cells
Anti-Idiotypic Antibodies
Salts
Binding Sites
Peptides
Ku Autoantigen
Antibodies

Keywords

  • human autoantigen Ku
  • transcription factor

ASJC Scopus subject areas

  • Genetics
  • General

Cite this

DNA-dependent protein kinase (Ku protein-p350 complex) assembles on double-stranded DNA. / Suwa, Akira; Hirakata, Michito; Takeda, Yoshihiko; Jesch, Stephen A.; Mimori, Tsuneyo; Hardin, John A.

In: Proceedings of the National Academy of Sciences of the United States of America, Vol. 91, No. 15, 19.07.1994, p. 6904-6908.

Research output: Contribution to journalArticle

Suwa, Akira ; Hirakata, Michito ; Takeda, Yoshihiko ; Jesch, Stephen A. ; Mimori, Tsuneyo ; Hardin, John A. / DNA-dependent protein kinase (Ku protein-p350 complex) assembles on double-stranded DNA. In: Proceedings of the National Academy of Sciences of the United States of America. 1994 ; Vol. 91, No. 15. pp. 6904-6908.
@article{84daabd7db6e4fedad7ec81a0f9dbaf1,
title = "DNA-dependent protein kinase (Ku protein-p350 complex) assembles on double-stranded DNA",
abstract = "The Ku protein is an autoantigen that consists of 70- and 80-kDa polypeptides. It associates with double-stranded DNA at free ends. In the present study, we examined the ability of anti-Ku antibodies to immunoprecipitate various structures from extracts of HeLa cells prepared at different salt concentrations. Under physiological conditions, these antibodies identified a complex containing the Ku protein and the 350-kDa component (p350) of DNA-dependent protein kinase (DNA-PK), which appeared to be closely associated on the DNA strand. In reconstitution experiments with cell extracts and biochemically purified components, the Ku protein-p350 complex formed only in the presence of double-stranded DNA. The reconstituted complex was catalytically active. Together with previous studies, these results indicate that the Ku protein interacts with DNA to create a binding site for p350 as the DNA-PK holoenzyme assembles.",
keywords = "human autoantigen Ku, transcription factor",
author = "Akira Suwa and Michito Hirakata and Yoshihiko Takeda and Jesch, {Stephen A.} and Tsuneyo Mimori and Hardin, {John A.}",
year = "1994",
month = "7",
day = "19",
doi = "10.1073/pnas.91.15.6904",
language = "English (US)",
volume = "91",
pages = "6904--6908",
journal = "Proceedings of the National Academy of Sciences of the United States of America",
issn = "0027-8424",
number = "15",

}

TY - JOUR

T1 - DNA-dependent protein kinase (Ku protein-p350 complex) assembles on double-stranded DNA

AU - Suwa, Akira

AU - Hirakata, Michito

AU - Takeda, Yoshihiko

AU - Jesch, Stephen A.

AU - Mimori, Tsuneyo

AU - Hardin, John A.

PY - 1994/7/19

Y1 - 1994/7/19

N2 - The Ku protein is an autoantigen that consists of 70- and 80-kDa polypeptides. It associates with double-stranded DNA at free ends. In the present study, we examined the ability of anti-Ku antibodies to immunoprecipitate various structures from extracts of HeLa cells prepared at different salt concentrations. Under physiological conditions, these antibodies identified a complex containing the Ku protein and the 350-kDa component (p350) of DNA-dependent protein kinase (DNA-PK), which appeared to be closely associated on the DNA strand. In reconstitution experiments with cell extracts and biochemically purified components, the Ku protein-p350 complex formed only in the presence of double-stranded DNA. The reconstituted complex was catalytically active. Together with previous studies, these results indicate that the Ku protein interacts with DNA to create a binding site for p350 as the DNA-PK holoenzyme assembles.

AB - The Ku protein is an autoantigen that consists of 70- and 80-kDa polypeptides. It associates with double-stranded DNA at free ends. In the present study, we examined the ability of anti-Ku antibodies to immunoprecipitate various structures from extracts of HeLa cells prepared at different salt concentrations. Under physiological conditions, these antibodies identified a complex containing the Ku protein and the 350-kDa component (p350) of DNA-dependent protein kinase (DNA-PK), which appeared to be closely associated on the DNA strand. In reconstitution experiments with cell extracts and biochemically purified components, the Ku protein-p350 complex formed only in the presence of double-stranded DNA. The reconstituted complex was catalytically active. Together with previous studies, these results indicate that the Ku protein interacts with DNA to create a binding site for p350 as the DNA-PK holoenzyme assembles.

KW - human autoantigen Ku

KW - transcription factor

UR - http://www.scopus.com/inward/record.url?scp=0028263785&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0028263785&partnerID=8YFLogxK

U2 - 10.1073/pnas.91.15.6904

DO - 10.1073/pnas.91.15.6904

M3 - Article

C2 - 8041718

AN - SCOPUS:0028263785

VL - 91

SP - 6904

EP - 6908

JO - Proceedings of the National Academy of Sciences of the United States of America

JF - Proceedings of the National Academy of Sciences of the United States of America

SN - 0027-8424

IS - 15

ER -