This Article
Right arrow Full Text
Right arrow Full Text (PDF)
Right arrow Supplemental material
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Services
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Download to citation manager
Right arrowReprints and Permissions
Right arrow Copyright Information
Right arrow Books from ASM Press
Right arrow MicrobeWorld
Citing Articles
Right arrow Citing Articles via HighWire
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Jiang, M.
Right arrow Articles by Maddock, J. R.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Jiang, M.
Right arrow Articles by Maddock, J. R.

 Previous Article  |  Next Article 

Journal of Bacteriology, May 2007, p. 3434-3444, Vol. 189, No. 9
0021-9193/07/$08.00+0     doi:10.1128/JB.00090-07
Copyright © 2007, American Society for Microbiology. All Rights Reserved.

Identification of Novel Escherichia coli Ribosome-Associated Proteins Using Isobaric Tags and Multidimensional Protein Identification Techniques{triangledown} ,{dagger}

M. Jiang,1 S. M. Sullivan,1 A. K. Walker,2 J. R. Strahler,2 P. C. Andrews,2 and J. R. Maddock1*

Department of Molecular, Cellular and Developmental Biology,1 National Resource for Proteomics and Pathways, University of Michigan, Ann Arbor, Michigan 481092

Received 17 January 2007/ Accepted 15 February 2007

Biogenesis of the large ribosomal subunit requires the coordinate assembly of two rRNAs and 33 ribosomal proteins. In vivo, additional ribosome assembly factors, such as helicases, GTPases, pseudouridine synthetases, and methyltransferases, are also critical for ribosome assembly. To identify novel ribosome-associated proteins, we used a proteomic approach (isotope tagging for relative and absolute quantitation) that allows for semiquantitation of proteins from complex protein mixtures. Ribosomal subunits were separated by sucrose density centrifugation, and the relevant fractions were pooled and analyzed. The utility and reproducibility of the technique were validated via a double duplex labeling method. Next, we examined proteins from 30S, 50S, and translating ribosomes isolated at both 16°C and 37°C. We show that the use of isobaric tags to quantify proteins from these particles is an excellent predictor of the particles with which the proteins associate. Moreover, in addition to bona fide ribosomal proteins, additional proteins that comigrated with different ribosomal particles were detected, including both known ribosomal assembly factors and unknown proteins. The ribosome association of several of these proteins, as well as others predicted to be associated with ribosomes, was verified by immunoblotting. Curiously, deletion mutants for the majority of these ribosome-associated proteins had little effect on cell growth or on the polyribosome profiles.

* Corresponding author. Mailing address: Department of Molecular, Cellular and Developmental Biology, University of Michigan, 830 North University, Ann Arbor, MI 48109-1048. Phone: (734) 936-8068. Fax: (734) 647-0884. E-mail: maddock{at}umich.edu

{triangledown} Published ahead of print on 2 March 2007.

{dagger} Supplemental material for this article may be found at http://jb.asm.org/.

Journal of Bacteriology, May 2007, p. 3434-3444, Vol. 189, No. 9
0021-9193/07/$08.00+0     doi:10.1128/JB.00090-07
Copyright © 2007, American Society for Microbiology. All Rights Reserved.



This article has been cited by other articles:

  • Keren, I., Klipcan, L., Bezawork-Geleta, A., Kolton, M., Shaya, F., Ostersetzer-Biran, O. (2008). Characterization of the Molecular Basis of Group II Intron RNA Recognition by CRS1-CRM Domains. J. Biol. Chem. 283: 23333-23342 [Abstract] [Full Text]  
  • DeVito, J. A. (2008). Recombineering with tolC as a Selectable/Counter-selectable Marker: remodeling the rRNA Operons of Escherichia coli. Nucleic Acids Res 36: e4-e4 [Abstract] [Full Text]  
  • Kaczanowska, M., Ryden-Aulin, M. (2007). Ribosome Biogenesis and the Translation Process in Escherichia coli. Microbiol. Mol. Biol. Rev. 71: 477-494 [Abstract] [Full Text]  
  • Jiang, M., Sullivan, S. M., Wout, P. K., Maddock, J. R. (2007). G-Protein Control of the Ribosome-Associated Stress Response Protein SpoT. J. Bacteriol. 189: 6140-6147 [Abstract] [Full Text]  


Copyright © 2009 by the American Society for Microbiology.   For an alternate route to Journals.ASM.org, visit: http://intl-journals.asm.org | More Info»