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Journal of Bacteriology, June 2002, p. 3260-3267, Vol. 184, No. 12
0021-9193/02/$04.00+0     DOI: 10.1128/JB.184.12.3260-3267.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.

In Vivo Analysis of an Essential Archaeal Signal Recognition Particle in Its Native Host

R. Wesley Rose and Mechthild Pohlschröder*

Department of Biology, University of Pennsylvania, Philadelphia, Pennsylvania 19104

Received 28 December 2001/ Accepted 15 March 2002

The evolutionarily conserved signal recognition particle (SRP) plays an integral role in Sec-mediated cotranslational protein translocation and membrane protein insertion, as it has been shown to target nascent secretory and membrane proteins to the bacterial and eukaryotic translocation pores. However, little is known about its function in archaea, since characterization of the SRP in this domain of life has thus far been limited to in vitro reconstitution studies of heterologously expressed archaeal SRP components identified by sequence comparisons. In the present study, the genes encoding the SRP54, SRP19, and 7S RNA homologs (hv54h, hv19h, and hv7Sh, respectively) of the genetically and biochemically tractable archaeon Haloferax volcanii were cloned, providing the tools to analyze the SRP in its native host. As part of this analysis, an hv54h knockout strain was created. In vivo characterization of this strain revealed that the archaeal SRP is required for viability, suggesting that cotranslational protein translocation is an essential process in archaea. Furthermore, a method for the purification of this SRP employing nickel chromatography was developed in H. volcanii, allowing the successful copurification of (i) Hv7Sh with a histidine-tagged Hv54h, as well as (ii) Hv54h and Hv7Sh with a histidine-tagged Hv19h. These results provide the first in vivo evidence that these components interact in archaea. Such copurification studies will provide insight into the significance of the similarities and differences of the protein-targeting systems of the three domains of life, thereby increasing knowledge about the recognition of translocated proteins in general.

* Corresponding author. Mailing address: Department of Biology, University of Pennsylvania, 201 Leidy Laboratories, 415 South University Ave., Philadelphia, PA 19104. Phone: (215) 573-2278. Fax: (215) 898-8780. E-mail: pohlschr{at}sas.upenn.edu.

Journal of Bacteriology, June 2002, p. 3260-3267, Vol. 184, No. 12
0021-9193/02/$04.00+0     DOI: 10.1128/JB.184.12.3260-3267.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.



This article has been cited by other articles:

  • Yurist, S., Dahan, I., Eichler, J. (2007). SRP19 Is a Dispensable Component of the Signal Recognition Particle in Archaea. J. Bacteriol. 189: 276-279 [Abstract] [Full Text]  
  • Hand, N. J., Klein, R., Laskewitz, A., Pohlschroder, M. (2006). Archaeal and Bacterial SecD and SecF Homologs Exhibit Striking Structural and Functional Conservation. J. Bacteriol. 188: 1251-1259 [Abstract] [Full Text]  
  • Haddad, A., Rose, R. W., Pohlschroder, M. (2005). The Haloferax volcanii FtsY Homolog Is Critical for Haloarchaeal Growth but Does Not Require the A Domain. J. Bacteriol. 187: 4015-4022 [Abstract] [Full Text]  
  • Ring, G., Eichler, J. (2004). In the Archaea Haloferax volcanii, Membrane Protein Biogenesis and Protein Synthesis Rates Are Affected by Decreased Ribosomal Binding to the Translocon. J. Biol. Chem. 279: 53160-53166 [Abstract] [Full Text]  
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