Skip to main content
  • ASM
    • Antimicrobial Agents and Chemotherapy
    • Applied and Environmental Microbiology
    • Clinical Microbiology Reviews
    • Clinical and Vaccine Immunology
    • EcoSal Plus
    • Eukaryotic Cell
    • Infection and Immunity
    • Journal of Bacteriology
    • Journal of Clinical Microbiology
    • Journal of Microbiology & Biology Education
    • Journal of Virology
    • mBio
    • Microbiology and Molecular Biology Reviews
    • Microbiology Resource Announcements
    • Microbiology Spectrum
    • Molecular and Cellular Biology
    • mSphere
    • mSystems
  • Log in
  • My alerts
  • My Cart

Main menu

  • Home
  • Articles
    • Current Issue
    • Accepted Manuscripts
    • Archive
    • Minireviews
  • For Authors
    • Submit a Manuscript
    • Scope
    • Editorial Policy
    • Submission, Review, & Publication Processes
    • Organization and Format
    • Errata, Author Corrections, Retractions
    • Illustrations and Tables
    • Nomenclature
    • Abbreviations and Conventions
    • Publication Fees
    • Ethics Resources and Policies
  • About the Journal
    • About MCB
    • Editor in Chief
    • Editorial Board
    • For Reviewers
    • For the Media
    • For Librarians
    • For Advertisers
    • Alerts
    • RSS
    • FAQ
  • Subscribe
    • Members
    • Institutions
  • ASM
    • Antimicrobial Agents and Chemotherapy
    • Applied and Environmental Microbiology
    • Clinical Microbiology Reviews
    • Clinical and Vaccine Immunology
    • EcoSal Plus
    • Eukaryotic Cell
    • Infection and Immunity
    • Journal of Bacteriology
    • Journal of Clinical Microbiology
    • Journal of Microbiology & Biology Education
    • Journal of Virology
    • mBio
    • Microbiology and Molecular Biology Reviews
    • Microbiology Resource Announcements
    • Microbiology Spectrum
    • Molecular and Cellular Biology
    • mSphere
    • mSystems

User menu

  • Log in
  • My alerts
  • My Cart

Search

  • Advanced search
Molecular and Cellular Biology
publisher-logosite-logo

Advanced Search

  • Home
  • Articles
    • Current Issue
    • Accepted Manuscripts
    • Archive
    • Minireviews
  • For Authors
    • Submit a Manuscript
    • Scope
    • Editorial Policy
    • Submission, Review, & Publication Processes
    • Organization and Format
    • Errata, Author Corrections, Retractions
    • Illustrations and Tables
    • Nomenclature
    • Abbreviations and Conventions
    • Publication Fees
    • Ethics Resources and Policies
  • About the Journal
    • About MCB
    • Editor in Chief
    • Editorial Board
    • For Reviewers
    • For the Media
    • For Librarians
    • For Advertisers
    • Alerts
    • RSS
    • FAQ
  • Subscribe
    • Members
    • Institutions
CELL GROWTH AND DEVELOPMENT

Proapoptotic p53-Interacting Protein 53BP2 Is Induced by UV Irradiation but Suppressed by p53

Charles D. Lopez, Yi Ao, Larry H. Rohde, Tomas D. Perez, Daniel J. O'Connor, Xin Lu, James M. Ford, Louie Naumovski
Charles D. Lopez
Divisions of Medical Oncology and
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Yi Ao
Division of Hematology/Oncology, Department of Pediatrics, Stanford University School of Medicine, Stanford, California 94305, and
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Larry H. Rohde
Division of Hematology/Oncology, Department of Pediatrics, Stanford University School of Medicine, Stanford, California 94305, and
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Tomas D. Perez
Division of Hematology/Oncology, Department of Pediatrics, Stanford University School of Medicine, Stanford, California 94305, and
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Daniel J. O'Connor
Ludwig Institute for Cancer Research, Imperial College of Medicine at St. Mary's, London W2 1PG, United Kingdom
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Xin Lu
Ludwig Institute for Cancer Research, Imperial College of Medicine at St. Mary's, London W2 1PG, United Kingdom
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
James M. Ford
Divisions of Medical Oncology and
Genetics, Department of Medicine, and
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Louie Naumovski
Division of Hematology/Oncology, Department of Pediatrics, Stanford University School of Medicine, Stanford, California 94305, and
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
DOI: 10.1128/MCB.20.21.8018-8025.2000
  • Article
  • Figures & Data
  • Info & Metrics
  • PDF
Loading

Article Figures & Data

Figures

  • Fig. 1.
    • Open in new tab
    • Download powerpoint
    Fig. 1.

    53BP2 levels increase following UV irradiation-induced DNA damage. (A) Western blots of lysates (30 μg of total protein per lane) prepared from GM38 cells (left panels) and HT1080 cells (right panels) 24 h after UV irradiation with 0, 5, or 10 J/m2. The fold increase of 53BP2 protein levels compared to no-UV-irradiation levels are indicated below, normalized for tubulin. 53BP2 was detected with monoclonal antibody DX547; p53 was detected with monoclonal antibody 1801. (B) Northern blot of total RNA (15 μg/lane) prepared from HT1080 cells 24 h after UV irradiation at the indicated doses, probed for 53BP2, stripped, and reprobed for actin. The fold increase of 53BP2 mRNA is indicated below, normalized for actin.

  • Fig. 2.
    • Open in new tab
    • Download powerpoint
    Fig. 2.

    53BP2 expression in cell lines containing wild-type or mutant p53. (A) The top panels show Western blots of lysates (30 μg of total protein per lane) prepared from human 041 and 087 LFS skin fibroblasts heterozygous for wild-type p53 (+/−) (lanes 2 and 4, respectively) or homozygous for p53 mutations (−/−) (lanes 3 and 5, respectively) and from normal primary human skin fibroblasts (GM38) homozygous for wild-type p53 (+/+) (lane 1). The fold differences, compared to GM38 cells, of 53BP2 protein normalized for tubulin are indicated below. 53BP2 protein was detected with monoclonal antibody DX547. The bottom panels show Northern blots of total RNA (15 μg/lane) prepared from cells as indicated above, probed for 53BP2, stripped, and reprobed for actin. The fold differences, compared to GM38 cells, of 53BP2 mRNA normalized for actin are indicated below. (B) Western blots of lysates (30 μg of total protein per lane) prepared from mouse fibroblasts homozygous for wild-type p53 (3T3, p21-null, and XPA+/−) or from p53 knockout mouse fibroblasts (p53-null). The fold differences, compared to 3T3 cells, of 53BP2 protein normalized for tubulin are indicated below. 53BP2 detected with rabbit antisera Rab-1.

  • Fig. 3.
    • Open in new tab
    • Download powerpoint
    Fig. 3.

    Expression of wild-type p53 in the absence of cellular damage decreases 53BP2 levels. (A) Western blot of lysates (30 μg of total protein per lane) prepared from 041TR cells at the indicated times after tetracycline withdrawal. The decreases in 53BP2 protein, as a fraction of baseline levels determined at time zero, are indicated below, normalized for tubulin. 53BP2 was detected with monoclonal antibody DX547; p53 was detected with monoclonal antibody 1801. (B) Northern blot of total RNA (15 μg/lane) prepared from the 041TR cells above at the indicated time points, probed for 53BP2, stripped, and reprobed for GAPDH. The decreases in 53BP2 mRNA, as a fraction of baseline levels determined at time zero, are indicated below, normalized for GAPDH.

  • Fig. 4.
    • Open in new tab
    • Download powerpoint
    Fig. 4.

    p53-independent mechanisms participate in the UV irradiation induction of 53BP2. (A) Western blot of lysates (15 μg of total protein per lane) prepared from 041−/− cells 24 h after UV irradiation with 0, 5, or 10 J/m2. The fold increase of 53BP2 protein compared to no-UV-irradiation lysates are indicated below, normalized for tubulin. (B) Western blot of equivalent amounts of lysates prepared from 041TR cells maintained in 2 μg of tetracycline per ml at the indicated times after UV irradiation at 20 J/m2. The fold increases of 53BP2 protein compared to the levels at time zero are indicated below, normalized for tubulin. (C) The upper panels show Western blots of equivalent amounts of lysates prepared from 041TR cells at the indicated times after tetracycline withdrawal. At time zero, cells were UV irradiated with 20 J/m2. The fold increases of 53BP2 protein compared to levels at time zero are indicated below, normalized for tubulin. 53BP2 protein was detected with monoclonal antibody DX547; p53 was detected with monoclonal antibody 1801. The lower panels show Northern blots of total RNA (15 μg/lane) prepared from the 041TR cells above at the indicated time points, probed for 53BP2, stripped, and reprobed for GAPDH. The fold increases of 53BP2 mRNA compared to the levels at time zero are indicated below, normalized for GAPDH.

  • Fig. 5.
    • Open in new tab
    • Download powerpoint
    Fig. 5.

    Expression of 53BP2 sensitizes cells to UV irradiation-induced apoptosis. (A) Percentage of 293/53BP2 and 293/LZ cells with apoptotic nuclear morphology 48 h after UV irradiation (shaded bars) or no UV irradiation (open bars). Cells induced to express 53BP2 or β-galactosidase (+) were treated with 2.5 μM ponasterone A for 24 h prior to UV irradiation. Mock-induced cells (−) were treated with an equivalent volume of control carrier (ethanol). Shown are the means and standard deviations from triplicate plates. (B) Western blot using anti-53BP2 monoclonal antibody DX547 of equivalent amounts of lysates prepared from 293/53BP2 cells induced for 24 h with 2.5 μM ponasterone A (+) or ethanol (−).

  • Fig. 6.
    • Open in new tab
    • Download powerpoint
    Fig. 6.

    Expression of 53BP2 decreases clonogenic survival in response to UV irradiation. (A) The left graph shows the surviving fraction of HT/53BP2 cells after induction of 53BP2 expression with 5 μM ponasterone A (■) or ethanol control carrier (⋄) for 48 h prior to UV irradiation at the indicated doses. Shown are the means and standard deviations from triplicate plates. The right graph shows the surviving fraction of HT/LZ cells after the induction of LacZ expression with 5 μM ponasterone A (■) or ethanol control carrier (⋄) for 48 h prior to UV irradiation at the indicated doses. Colonies were fixed, stained, and counted after 12 days. (B) Western blot using anti-53BP2 monoclonal antibody DX547 of equivalent amounts of lysates prepared from HT/53BP2 cells induced for 48 h with 5 μM ponasterone A (+) or ethanol (−).

  • Fig. 7.
    • Open in new tab
    • Download powerpoint
    Fig. 7.

    Attenuation of 53BP2 induction enhances clonogenic survival following UV irradiation. (A) Surviving fraction of HT1080 cells after introduction of AS-oligo (■) or control-oligo (⋄). Oligonucleotide-treated cells were incubated for 5 h, followed by counting, replating, and UV irradiation at the indicated doses. Shown is a representative experiment with the means and standard deviations from triplicate plates. (B) Western blot using anti-53BP2 monoclonal antibody DX547 of equivalent amounts of lysates (50 μg of total protein per lane) prepared from HT1080 cells after the introduction of AS-oligo or control-oligo. Oligonucleotide treated cells were incubated and UV irradiated in parallel with the clonogenic assay. Lysates were prepared 24 h after UV irradiation. Anti-hsc70 monoclonal antibody was used to confirm equal loading.

  • Fig. 8.
    • Open in new tab
    • Download powerpoint
    Fig. 8.

    Model of 53BP2 pathways. In undamaged cells, p53 mediates the suppression of 53BP2 levels. UV damage can relieve this suppression and increase 53BP2 levels. Through unknown mechanisms, 53BP2 can inhibit cell growth and promote damage-induced apoptosis after UV irradiation, as well as enhance p53 transactivation and cell cycle arrest. Lines with arrowheads denote positive regulation; the slashed line denotes negative regulation. “p53*” denotes the DNA damage-activated state.

PreviousNext
Back to top
Download PDF
Citation Tools
Proapoptotic p53-Interacting Protein 53BP2 Is Induced by UV Irradiation but Suppressed by p53
Charles D. Lopez, Yi Ao, Larry H. Rohde, Tomas D. Perez, Daniel J. O'Connor, Xin Lu, James M. Ford, Louie Naumovski
Molecular and Cellular Biology Nov 2000, 20 (21) 8018-8025; DOI: 10.1128/MCB.20.21.8018-8025.2000

Citation Manager Formats

  • BibTeX
  • Bookends
  • EasyBib
  • EndNote (tagged)
  • EndNote 8 (xml)
  • Medlars
  • Mendeley
  • Papers
  • RefWorks Tagged
  • Ref Manager
  • RIS
  • Zotero
Print

Alerts
Sign In to Email Alerts with your Email Address
Email

Thank you for sharing this Molecular and Cellular Biology article.

NOTE: We request your email address only to inform the recipient that it was you who recommended this article, and that it is not junk mail. We do not retain these email addresses.

Enter multiple addresses on separate lines or separate them with commas.
Proapoptotic p53-Interacting Protein 53BP2 Is Induced by UV Irradiation but Suppressed by p53
(Your Name) has forwarded a page to you from Molecular and Cellular Biology
(Your Name) thought you would be interested in this article in Molecular and Cellular Biology.
CAPTCHA
This question is for testing whether or not you are a human visitor and to prevent automated spam submissions.
Share
Proapoptotic p53-Interacting Protein 53BP2 Is Induced by UV Irradiation but Suppressed by p53
Charles D. Lopez, Yi Ao, Larry H. Rohde, Tomas D. Perez, Daniel J. O'Connor, Xin Lu, James M. Ford, Louie Naumovski
Molecular and Cellular Biology Nov 2000, 20 (21) 8018-8025; DOI: 10.1128/MCB.20.21.8018-8025.2000
del.icio.us logo Digg logo Reddit logo Twitter logo CiteULike logo Facebook logo Google logo Mendeley logo
  • Top
  • Article
    • ABSTRACT
    • MATERIALS AND METHODS
    • RESULTS
    • DISCUSSION
    • ACKNOWLEDGMENTS
    • FOOTNOTES
    • REFERENCES
  • Figures & Data
  • Info & Metrics
  • PDF

Related Articles

Cited By...

About

  • About MCB
  • Editor in Chief
  • Editorial Board
  • Policies
  • For Reviewers
  • For the Media
  • For Librarians
  • For Advertisers
  • Alerts
  • RSS
  • FAQ
  • Permissions
  • Journal Announcements

Authors

  • ASM Author Center
  • Submit a Manuscript
  • Article Types
  • Ethics
  • Contact Us

Follow #MCBJournal

@ASMicrobiology

       

ASM Journals

ASM journals are the most prominent publications in the field, delivering up-to-date and authoritative coverage of both basic and clinical microbiology.

About ASM | Contact Us | Press Room

 

ASM is a member of

Scientific Society Publisher Alliance

 

American Society for Microbiology
1752 N St. NW
Washington, DC 20036
Phone: (202) 737-3600

Copyright © 2021 American Society for Microbiology | Privacy Policy | Website feedback

Print ISSN: 0270-7306; Online ISSN: 1098-5549