Skip to main content
SpringerLink
Log in

Mutations in PurBox1 of theBacillus subtilis pur operon control site affect adenine-regulated expressionin vivo

  • Published:
Science in China Series C: Life Sciences Aims and scope Submit manuscript

Abstract

Transcription of theBacillus subtilis pur operon is regulated by a purine repressor (PurR)-DNA control site interaction. Thepur operon control site has two PurBoxes that are required for high-affinity PurR binding. An upstream, strong-binding PurBox1 is at position −81 to −68 relative to the transcription start site and a downstream weak-binding PurBox2 is at position −49 to −36. We constructed three PurBox1 mutations and the effects on binding of PurR to the control regionin vitro and on regulation ofpur operon expressionin vivo were investigated. The mutations significantly reduced the binding of PurR to control region DNA. In strains with G-75A, G-75T and a five bp deletion (Δ5)pur operon repression was defectivein vivo. In additionin vivo PurR titration was used to confirm that sequences flanking PurBox1 and PurBox2 are required for PurR binding to thepur operon control site.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

References

  1. Ebbole, D. J., Zalkin, H., Cloning and characterization of a 12-gene cluster fromBacillus subtilis encoding nine enzymes forde novo purine nucleotide synthesis, J. Biol. Chem., 1987, 262(17): 8274–8287.

    PubMed  CAS  Google Scholar 

  2. Ebbole, D. J., Zalkin, H., Detection of pur operon attenuated mRNA and accumulated degradation intermediates inBacillus subtilis, J. Biol. Chem., 1988, 263(22): 10894–10902.

    PubMed  CAS  Google Scholar 

  3. Ebbole, D. J., Zalkin, H.,Bacillus subtilis pur operon expression and regulation, J. Bacteriol., 1989, 171(4): 2136–2141.

    PubMed  CAS  Google Scholar 

  4. Ebbole, D. J., Zalkin, H., Interaction of a putative repressor protein with an extended control region of theBacillus subtilis pur operon, J. Biol. Chem., 1989, 264(6): 3553–3561.

    PubMed  CAS  Google Scholar 

  5. Weng, M., Nagy, P. L., Zalkin, H., Identification of theBacillus subtilis pur operon repressor, Proc. Natl. Acad. Sci. USA, 1995, 92: 7455–7459.

    Article  PubMed  CAS  Google Scholar 

  6. Weng, M., Zalkin, H., Mutations in theBacillus subtilis purine repressor that perturb PRPP effector functionin vitro andin vivo, Current Microbiol., 2000, 41: 56–59.

    Article  PubMed  CAS  Google Scholar 

  7. Shin, B. S., Stein, A., Zalkin, H., Interaction ofBacillus subtilis purine repressor with DNA, J. Bacteriol., 1997, 179(23): 7394–7402.

    PubMed  CAS  Google Scholar 

  8. Bera, A. K., Zhu, J., Zalkin, H., Smith, J. L., Functional dissection of theBacillus subtilis purine operator site, J. Bacteriol., 2003, 185(14): 4099–4109.

    Article  PubMed  CAS  Google Scholar 

  9. Rappu, P., Shin, B. S., Zalkin, H., Mantsala, P., A role for a highly conserved protein of unknown function in regulation ofBacillus subtilis purA by the purine repressor, J. Bacteriol., 1999, 181(12): 3810–3815.

    PubMed  CAS  Google Scholar 

  10. Kilstrup, M., Martinussen, J., A transcriptional activator, homologous to theBacillus subtilis PurR repressor, is required for expression of purine biosynthetic genes inLactococcus lactis, J. Bacteriol., 1998, 180(15): 3907–3916.

    PubMed  CAS  Google Scholar 

  11. Kilstrup, M., Jessing, S. G., Wichmand-Jorgensen, S., Madsen, M., Nilsson, D., Activation control of pur gene expression inLactococcus lactis: Proposal for a consensus activator binding sequence based on deletion analysis and site-directed mutagenesis of purC and purD promoter regions, J. Bacteriol., 1998, 180(15): 3900–3906.

    PubMed  CAS  Google Scholar 

  12. Bruckner, R., A series of shuttle vectors forBacillus subtilis andEscherichia coli, Gene, 1992, 122: 187–192.

    Article  PubMed  CAS  Google Scholar 

  13. Miller, J. H., A short course in bacterial genetics. Plainview, NY: Cold Spring Harber Laboratory Press, 1992, 72–79, 437–439.

    Google Scholar 

  14. Bradford, M. M., A rapid and sensitive for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding, Anal. Biochem., 1976, 72: 248–254.

    Article  PubMed  CAS  Google Scholar 

  15. Kuroda, M. I., Henner, D., Yanofsky, C., Cis-acting sites in the transcript of theBacillus subtilis trp operon regulate expression of the operon, J. Bacteriol., 1988, 170(7): 3080–3088.

    PubMed  CAS  Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Institute of Genetics and Developmental Biology, Chinese Academy of Sciences, 100101, Beijing, China

    Jinsong Xuan & Manli Weng

  2. Department of Biochemistry, Purdue University, IN 47907, West Lafayette, USA

    Howard Zalkin

Authors
  1. Jinsong Xuan
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Howard Zalkin
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Manli Weng
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Manli Weng.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Xuan, J., Zalkin, H. & Weng, M. Mutations in PurBox1 of theBacillus subtilis pur operon control site affect adenine-regulated expressionin vivo . Sci. China Ser. C.-Life Sci. 48, 133–138 (2005). https://doi.org/10.1007/BF02879665

Download citation

  • Received:

  • Revised:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF02879665

Keywords

Search

Navigation