FCCC LOGO Faculty Publications
Haddad A , Rose RW , Pohlschro?der M
The Haloferax volcanii FtsY homolog is critical for haloarchaeal growth but does not require the A domain
Journal of Bacteriology. 2005 ;187(12) :4015-4022
Back to previous list
The targeting of many Sec substrates to the membrane-associated translocation pore requires the cytoplasmic signal recognition particle (SRP). In Eukarya and Bacteria it has been shown that membrane docking of the SRP-substrate complex occurs via the universally conserved SRP receptor (Sr?/? and FtsY, respectively). While much has been learned about the archaeal SRP in recent years, few studies have examined archaeal Sr?/FtsY homologs. In the present study the FtsY homolog of Haloferax volcanii was characterized in its native host. Disruption of the sole chromosomal copy of ftsY in H. volcanii was possible only under conditions where either the full-length haloarchaeal FtsY or an amino-terminally truncated version of this protein lacking the A domain, was expressed in trans. Subcellular fractionation analysis of H. volcanii ftsY deletion strains expressing either one of the complementing proteins revealed that in addition to a cytoplasmic pool, both proteins cofractionate with the haloarchaeal cytoplasmic membrane. Moreover, membrane localization of the universally conserved SRP subunit SRP54, the key binding partner of FtsY, was detected in both H. volcanii strains. These analyses suggest that the H. volcanii FtsY homolog plays a crucial role but does not require its A domain for haloarchaeal growth. Copyright © 2005, American Society for Microbiology. All Rights Reserved.
00219193 (ISSN) Cited By: 3; Export Date: 25 May 2006; Source: Scopus CODEN: JOBAA; DOI: 10.1128/JB.187.12.4015-4022.2005 Language of Original Document: English Correspondence Address: Pohlschro?der, M.; Department of Biology; University of Pennsylvania; 201 Leidy Laboratories; 415 South University Ave. Philadelphia, PA 19104, United States; email: pohlschr@sas.upenn.edu Molecular Sequence Numbers: GENBANK: AY187867; Chemicals/CAS: Bacterial Proteins; FtsY protein, Bacteria; Receptors, Cytoplasmic and Nuclear References: Allers, T., Ngo, H.P., Mevarech, M., Lloyd, R.G., Development of additional selectable markers for the halophilic archaeon Haloferax volcanii based on the leuB and trpA genes (2004) Appl. Environ. Microbiol., 70, pp. 943-953; Bitan-Banin, G., Ortenberg, R., Mevarech, M., Development of a gene knockout system for the halophilic archaeon Haloferax volcanii by use of the pyrE gene (2003) J. Bacteriol., 185, pp. 772-778; Bonneau, R., Baliga, N.S., Deutsch, E.W., Shannon, P., Hood, L., Comprehensive de novo structure prediction in a systems-biology context for the archaea Halobacterium sp. NRC-1 (2004) Genome Biol., 5, pp. R52; Charlebois, R.L., Lam, W.L., Cline, S.W., Doolittle, W.F., Characterization of pHV2 from Halobacterium volcanii and its use in demonstrating transformation of a archaebacterium (1987) Proc. Natl. Acad. Sci. USA, 84, pp. 8530-8534; Charlebois, R.L., Schalkwyk, L.C., Hofman, J.D., Doolittle, W.F., Detailed physical map and set of overlapping clones covering the genome of the archaebacterium Haloferax volcanii DS2 (1991) J. Mol. Biol., 222, pp. 509-524; De Leeuw, E., Poland, D., Mol, O., Sinning, I., Ten Hagen-Jongman, C.M., Oudega, B., Luirink, J., Membrane association of FtSY, the E. coli SRP receptor (1997) FEBS Lett., 416, pp. 225-229; De Leeuw, E., Te Kaat, K., Moser, C., Menestrina, G., Demel, R., De Kruijff, B., Oudega, B., (...), Sinning, I., Anionic phospholipids are involved in membrane association of FtsY and stimulate its GTPase activity (2000) EMBO J., 19, pp. 531-541; Doudna, J.A., Batey, R.T., Structural insights into the signal recognition particle (2004) Annu. Rev. Biochem., 73, pp. 539-557; Egea, P.F., Shan, S.O., Napetschnig, J., Savage, D.F., Walter, P., Stroud, R.M., Substrate twinning activates the signal recognition particle and its receptor (2004) Nature, 427, pp. 215-221; Eichler, J., Archaeal protein translocation: Crossing membranes in the third domain of life (2000) Eur. J. Biochem., 267, pp. 3402-3412; Eitan, A., Bibi, E., The core Escherichia coli signal recognition particle receptor contains only the N and G domains of FtsY (2004) J. Bacteriol., 186, pp. 2492-2494; Focia, P.J., Shepotinovskaya, I.V., Seidler, J.A., Freymann, D.M., Heterodimeric GTPase core of the SRP targeting complex (2004) Science, 303, pp. 373-377; Fulga, T.A., Sinning, I., Dobberstein, B., Pool, M.R., SR? coordinates signal sequence release from SRP with ribosome binding to the translocon (2001) EMBO J., 20, pp. 2338-2347; Futterer, O., Angelov, A., Liesegang, H., Gottschalk, G., Schleper, C., Schepers, B., Dock, C., (...), Liebl, W., Genome sequence of Picrophilus torridus and its implications for life around pH 0 (2004) Proc. Natl. Acad. Sci. USA, 101, pp. 9091-9096; Gill, D.R., Salmond, G.P.C., The identification of the Escherichia coli ftsY gene product: An unusual protein (1990) Mol. Microbiol., 4, pp. 575-583; Gropp, R., Gropp, F., Betlach, M.C., Association of the halobacterial 7S RNA to the polysome correlates with expression of the membrane protein bacterioopsin (1992) Proc. Natl. Acad. Sci. USA, 89, pp. 1204-1208; Jolley, K.A., Rapaport, E., Hough, D.W., Danson, M.J., Woods, W.G., Dyall-Smith, M.L., Dihydrolipoamide dehydrogenase from the halophilic archaeon Haloferax volcanii: Homologous overexpression of the cloned gene (1996) J. Bacteriol., 178, pp. 3044-3048; Keeling, P.J., Charlebois, R.L., Doolittle, W.F., Archaebacterial genomes: Eubacterial form and eukaryotic content (1994) Curr. Opin. Gene Dev., 4, pp. 816-833; Lichi, T., Ring, G., Eichler, J., Membrane binding of SRP pathway components in the halophilic archaea Haloferax volcanii (2004) Eur. J. Biochem., 271, pp. 1382-1390; Luirink, J., Ten Hagen-Jongman, C.M., Van Der Weijden, C.C., Oudega, B., High, S., Dobberstein, B., Kusters, R., An alternative protein targeting pathway in Escherichia coli: Studies on the role of FtsY (1994) EMBO J., 13, pp. 2289-2296; Lund, P.A., Large, A.T., Kapatai, G., The chaperonins: Perspectives from the Archaea (2003) Biochem. Soc. Trans., 31, pp. 681-685; Mevarech, M., Werczberger, R., Genetic transfer in Halobacterium volcanii (1985) J. Bacteriol., 162, pp. 461-462; Millman, J.S., Andrews, D.W., A site-specific, membrane-dependent cleavage event defines the membrane binding domain of FtsY (1999) J. Biol. Chem., 274, pp. 33227-33234; Millman, J.S., Qi, H.Y., Vulcu, F., Bernstein, H.D., Andrews, D.W., FtsY binds to the Escherichia coli inner membrane via interactions with phosphatidylethanolamine and membrane proteins (2001) J. Biol. Chem., 276, pp. 25982-25989; Moll, R.G., Protein-protein, protein-RNA and protein-lipid interactions of signal-recognition particle components in the hyperthermoacidophilic archaeon Acidianus ambivalens (2003) Biochem. J., 374, pp. 247-254; Ogg, S.C., Barz, W.P., Walter, P., A functional GTPase domain, but not its transmembrane domain, is required for function of the SRP receptor beta-subunit (1998) J. Cell Biol., 142, pp. 341-354; Patenge, N., Soppa, J., Extensive proteolysis inhibits high-level production of eukaryal G protein-coupled receptors in the archaeon Haloferax volcanii (1999) FEMS Microbiol. Lett., 171, pp. 27-35; Pogliano, K.J., Beckwith, J., The Cs sec mutants of Escherichia coli reflect the cold sensitivity of protein export itself (1993) Genetics, 133, pp. 763-773; Pohlschro?der, M., Dilks, K., Hand, N.J., Wesley Rose, R., Translocation of proteins across archaeal cytoplasmic membranes (2004) FEMS Microbiol. Rev., 28, pp. 3-24; Pohlschro?der, M., Murphy, C., Beckwith, J., In vivo analyses of interactions between SecE and SecY, core components of the Escherichia coli protein translocation machinery (1996) J. Biol. Chem., 271, pp. 19908-19914; Pohlschro?der, M., Prinz, W.A., Hartmann, E., Beckwith, J., Protein translocation in the three domains of life: Variations on a theme (1997) Cell, 91, pp. 563-566; Pool, M.R., Getting to the membrane: How is co-translational protein targeting to the endoplasmic reticulum regulated? (2003) Biochem. Soc. Trans., 31, pp. 1232-1237; Ring, G., Eichler, J., In the Archaea Haloferax volcanii, membrane protein biogenesis and protein synthesis rates are affected by decreased ribosomal binding to the translocon (2004) J. Biol. Chem., 279, pp. 53160-53166; Robinson, N.P., Dionne, I., Lundgren, M., Marsh, V.L., Bernander, R., Bell, S.D., Identification of two origins of replication in the single chromosome of the archaeon Sulfolobus solfataricus (2004) Cell, 116, pp. 25-38; Rose, R.W., Pohlschro?der, M., In vivo analysis of an essential archaeal signal recognition particle in its native host (2002) J. Bacteriol., 184, pp. 3260-3267; Sambrook, J., Fritsch, E.F., Maniatis, T., (1989) Molecular Cloning: A Laboratory Manual, 1, Cold Spring Harbor Laboratory Press. Cold Spring Harbor, N.Y.; Schut, G.J., Brehm, S.D., Datta, S., Adams, M.W., Whole-genome DNA microarray analysis of a hyperthermophile and an archaeon: Pyrococcus furiosus grown on carbohydrates or peptides (2003) J. Bacteriol., 185, pp. 3935-3947; Slesarev, A.I., Mezhevaya, K.V., Makarova, K.S., Polushin, N.N., Shcherbinina, O.V., Shakhova, V.V., Belova, G.I., (...), Kozyavkin, S.A., The complete genome of hyperthermophile Methanopyrus kandleri AV19 and monophyly of archaeal methanogens (2002) Proc. Natl. Acad. Sci. USA, 99, pp. 4644-4649; Tian, H., Beckwith, J., Genetic screen yields mutations in genes encoding all known components of the Escherichia coli signal recognition particle pathway (2002) J. Bacteriol., 184, pp. 111-118; Tian, H., Boyd, D., Beckwith, J., A mutant hunt for defects in membrane protein assembly yields mutations affecting the bacterial signal recognition particle and Sec machinery (2000) Proc. Natl. Acad. Sci. USA, 97, pp. 4730-4735; Tozik, I., Huang, Q., Zwieb, C., Eichler, J., Reconstitution of the signal recognition particle of the halophilic archaeon Haloferax volcanii (2002) Nucleic Acids Res., 30, pp. 4166-4175; Van Den Berg, B., Demons Jr., W.M., Collinson, I., Modis, Y., Hartmann, E., Harrison, S.C., Rapoport, T.A., X-ray structure of a protein-conducting channel (2004) Nature, 427, pp. 36-44; Walter, P., Blobel, G., Purification of a membrane-associated protein complex required for protein translocation across the endoplasmic reticulum (1980) Proc. Natl. Acad. Sci. USA, 77, pp. 7112-7116; Wild, K., Halic, M., Sinning, I., Beckmann, R., SRP meets the ribosome (2004) Nat. Struct. Mol. Biol., 11, pp. 1049-1053; Woese, C.R., Kandler, O., Wheelis, M.L., Toward a natural system of organisms: Proposal for the domains Archaca, Bacteria, and Eukarya (1990) Proc. Natl. Acad. Sci. USA, 87, pp. 4576-4579.