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 Cart

Main menu

  • Home
  • Articles
    • Archive
  • About the Journal
    • About EC
    • For Librarians
    • For Advertisers
    • FAQ
  • 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 Cart

Search

  • Advanced search
Eukaryotic Cell
publisher-logosite-logo

Advanced Search

  • Home
  • Articles
    • Archive
  • About the Journal
    • About EC
    • For Librarians
    • For Advertisers
    • FAQ
Articles

Bul Proteins, a Nonredundant, Antagonistic Family of Ubiquitin Ligase Regulatory Proteins

Tatiana V. Novoselova, Kiran Zahira, Ruth-Sarah Rose, James A. Sullivan
Tatiana V. Novoselova
School of Biological and Chemical Sciences, Queen Mary, University of London, London, United Kingdom
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Kiran Zahira
School of Biological and Chemical Sciences, Queen Mary, University of London, London, United Kingdom
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Ruth-Sarah Rose
School of Biological and Chemical Sciences, Queen Mary, University of London, London, United Kingdom
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
James A. Sullivan
School of Biological and Chemical Sciences, Queen Mary, University of London, London, United Kingdom
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
DOI: 10.1128/EC.00009-12
  • Article
  • Figures & Data
  • Info & Metrics
  • PDF
Loading

Article Figures & Data

Figures

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

    BUL genes are not essential for viability but are important for response to heat stress. (A) HA epitope-tagged Bul3p was expressed in a bul3Δ null mutant strain, and the protein was visualized by immunoblotting using anti-HA antibodies. The arrowheads indicate two major translation products of 55 to 60 kDa; the asterisk indicates a readthrough translation product of ∼100 kDa. (B) HABul3p was expressed in a bul3Δ strain, along with a Y100A (HABul3ΔPY) form, a variant that lacks the internal stop codon (HABul3ns), and an empty HA vector control. Extracts from log-phase cells (Input) were subjected to immunoprecipitation with anti-HA affinity resin (IP αHA) and subjected to immunoblotting with anti-HA and anti-Rsp5 antibodies. (C) Tenfold serial dilution of yeast strains with the BUL genes deleted alone (Δ1, Δ2, and Δ3), in pairs (Δ1,2; Δ1,3; and Δ2,3), or in combination (Δ1,2,3) were grown for 3 days at either 30°C or 42°C. WT, wild type. (D) Serial deletions as described for panel C with strains expressing HABul3 or HABul3ΔPY constructs. (E and F) In vitro ubiquitination assays using recombinant S-tagged Bul1p and Bul2p and large (Bul3ns) and small (Bul3stop) forms of Bul3p. The reactions were performed with methylated ubiquitin (Ub) using equal quantities, as determined using immunoblots with anti-Rsp5p antibodies (F), of wild-type Rsp5p and variants that had only a single functional WW domain remaining (WW3, WW2, and WW1) before being subjected to immunoblotting with anti-S-tag antibodies.

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

    BUL gene deletions show complex growth phenotypes but are not required for ubiquitin-mediated processing of Mga2p. (A) Tenfold serial dilution of BUL gene deletion strains grown on YPD agar medium, YPD plus 25 μM CdCl2 (Cd) or 3 μM phleomycin (Phl), or YEP plus 2% (vol/vol) ethanol (EtOH). (B) Tenfold serial dilution of BUL gene deletion strains grown on YPD agar medium plus 1 mM AZC, synthetic complete medium with no arginine (SC), or SC minus Arg plus 14 μM canavanine (Can). (C) Immunoblot using anti-HA antibodies of HA-Mga2p processing from P120 to P90 in wild-type and BUL gene deletion (Δ1,2,3) strains.

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

    Sorting of mCherryΔNSmf1p in metal-depleted medium following the addition of CdCl2. (A) mCherry fluorescence and differential interference contrast microscopy images of wild-type and single BUL gene deletion strains expressing mCherryΔNSmf1p following growth on metal-depleted media (−metal) and 2 h after the addition of 25 μM CdCl2. The arrowheads indicate plasma membrane localization. (B) (Top) Immunoblots against Tap-tagged ΔNSmf1p in total protein extracts from wild-type and bul1Δ (Δ1) and bul2Δ (Δ2) strains. (Bottom) PonceauS staining was used as a loading control.

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

    Sorting of GFPGap1p in nitrogen-poor medium. Shown are GFP fluorescence and differential interference contrast microscopy images of wild-type and single BUL gene deletion strains expressing GFPGap1p following growth in medium with proline as the sole nitrogen source.

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

    Sorting of GFPCan1p following growth in excess arginine. Shown are GFP fluorescence and differential interference contrast microscopy images of wild-type and single BUL gene deletion strains expressing GFPCan1p. The images were obtained before (−Arg) and 2 h after (+Arg) the addition of 100 mg/liter arginine to log-phase cells grown in arginine-free synthetic complete medium. The arrowheads indicate plasma membrane localization.

Tables

  • Figures
  • Table 1

    Percent identity/similarity of the Bul proteinsa

    Protein% Identity/similarity to:
    Bul1Bul2Bul3
    Bul151/6631/50
    Bul251/6630/51
    Bul331/5030/51
    • ↵a The table compares the global percent amino acid identity and similarity of Bul1p, Bul2p, and Bul3p (including the bypass stop codon).

PreviousNext
Back to top
Download PDF
Citation Tools
Bul Proteins, a Nonredundant, Antagonistic Family of Ubiquitin Ligase Regulatory Proteins
Tatiana V. Novoselova, Kiran Zahira, Ruth-Sarah Rose, James A. Sullivan
Eukaryotic Cell Mar 2012, 11 (4) 463-470; DOI: 10.1128/EC.00009-12

Citation Manager Formats

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

Email

Thank you for sharing this Eukaryotic Cell 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.
Bul Proteins, a Nonredundant, Antagonistic Family of Ubiquitin Ligase Regulatory Proteins
(Your Name) has forwarded a page to you from Eukaryotic Cell
(Your Name) thought you would be interested in this article in Eukaryotic Cell.
CAPTCHA
This question is for testing whether or not you are a human visitor and to prevent automated spam submissions.
Share
Bul Proteins, a Nonredundant, Antagonistic Family of Ubiquitin Ligase Regulatory Proteins
Tatiana V. Novoselova, Kiran Zahira, Ruth-Sarah Rose, James A. Sullivan
Eukaryotic Cell Mar 2012, 11 (4) 463-470; DOI: 10.1128/EC.00009-12
del.icio.us logo Digg logo Reddit logo Twitter logo CiteULike logo Facebook logo Google logo Mendeley logo
  • Top
  • Article
    • ABSTRACT
    • INTRODUCTION
    • MATERIALS AND METHODS
    • RESULTS
    • DISCUSSION
    • ACKNOWLEDGMENTS
    • FOOTNOTES
    • REFERENCES
  • Figures & Data
  • Info & Metrics
  • PDF

Related Articles

Cited By...

About

  • About EC
  • For the Media
  • For Librarians
  • For Advertisers
  • FAQ
  • Permissions
  • Journal Announcements

Authors

  • Submit a Manuscript to mSphere

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: 1535-9778; Online ISSN: 1535-9786