Skip to main content
SpringerLink
Log in

Genetic Studies on Protein Export in Bacteria

  • Conference paper
Protein Secretion and Export in Bacteria

Part of the book series: Current Topics in Microbiology and Immunology ((CT MICROBIOLOGY,volume 125))

Abstract

The study of protein export has proceeded in parallel in eukaryotic and prokaryotic systems over the past 10 years. Much of the information on eukaryotic protein secretion has come from the detailed analysis of an in vitro system which appears faithfully to mimic the in vivo pathway. In prokaryotes much of the progress has come from in vivo analysis including genetic studies. The genetic studies have focussed both on the components of specific proteins which determine their localization and on the nature of cellular components which are part of the export process. There now exists a large collection of mutations in the signal sequences of bacterial envelope proteins. These have been accumulated using both in vivo selections and in vitro constructions. These mutations have confirmed many aspects of the original signal sequence hypothesis and have provided some insight into the important features of such sequences. Studies on signal sequence mutations have been recently reviewed (Michaelis and Beckwith 1982; Benson et al. 1985) and will not be covered in this article.

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

Access this chapter

Log in via an institution
Chapter
USD 29.95
Price excludes VAT (USA)
  • Available as PDF
  • Read on any device
  • Instant download
  • Own it forever
eBook
USD 84.99
Price excludes VAT (USA)
  • Available as PDF
  • Read on any device
  • Instant download
  • Own it forever
Softcover Book
USD 109.99
Price excludes VAT (USA)
  • Compact, lightweight edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info

Tax calculation will be finalised at checkout

Purchases are for personal use only

Institutional subscriptions

Preview

Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.

Similar content being viewed by others

References

  • Bankaitis VA, Bassford PJ Jr (1985) Proper interaction between at least two components is required for efficient export of proteins to the Escherichia coli. J Bacteriol 161: 169–178

    PubMed  CAS  Google Scholar 

  • Bassford P, Beckwith J (1979) Mutants of Escherichia coli which accumulate the precursor of a secreted protein in the cytoplasm. Nature 277: 538–541

    Article  PubMed  CAS  Google Scholar 

  • Bassford P, Silhavy T, Beckwith J (1979) Use of gene fusion to study secretion of maltose-binding protein in Escherichia coli periplasm. J Bacteriol 139: 19–31

    PubMed  CAS  Google Scholar 

  • Beacham IR (1979) Periplasmic enzymes in gram-negative bacteria. Int J Biochem 10: 877–883

    Article  PubMed  CAS  Google Scholar 

  • Beckwith JR, Galizzi A, Smith GR (1985) The tools of bacterial genetics. In: Galizzi A, Scaife J (eds) Genetics of bacteria. Academic, London, pp 1–23

    Google Scholar 

  • Benson SA, Bremer E, Silhavy TJ (1984) Intragenic region required for LamB export. Proc Natl Acad Sci USA 81: 3830–3834

    Article  PubMed  CAS  Google Scholar 

  • Benson SA, Hall MN, Silhavy TJ (1985) Genetic analysis of protein export in Escherichia coli K-12. Annu Rev Biochem 54: 101–134

    Article  PubMed  CAS  Google Scholar 

  • Boeke JD, Model P (1982) A prokaryotic membrane anchor sequence: carboxyl terminus of bacteriophage f1 gene III protein retains it in the membrane. Proc Natl Acad Sci USA 79: 5200–5204

    Article  PubMed  CAS  Google Scholar 

  • Brickman ER, Oliver DB, Garwin JL, Kumamoto C (1984) The use of extragenic suppressors to define genes involved in protein export in Escherichia coli. MGG 196: 24–27

    PubMed  CAS  Google Scholar 

  • Cerretti DP, Dean D, Davis GR, Bedwell DM, Nomura M (1983) The spc ribosomal protein operon of Escherichia coli: sequence and cotranscription of the ribosomal protein genes and a protein export gene. Nucleic Acids Res 11 -. 2599–2616

    Google Scholar 

  • Chan SJ, Weiss J, Konrad M, White T, Bahl C, Yu S-D, Marks D, Steiner DF (1981) Biosynthesis and periplasmic segregation of human proinsulin in Escherichia coli. Proc Natl Acad Sci USA 78: 5401–5405

    Article  PubMed  CAS  Google Scholar 

  • Chen L, Rhoads D, Tai PC (1985) Alkaline phosphatase and OmpA protein can be translocated posttranslationally into membrane vesicles of Escherichia coli. J Bacteriol 161: 973–980

    PubMed  CAS  Google Scholar 

  • Emr SD, Bassford PJ Jr (1982) Localization and processing of outer membrane and periplasmic proteins in Escherichia coli strains harboring export-specific suppressor mutations. J Biol Chem 257: 5852–5860

    PubMed  CAS  Google Scholar 

  • Emr SD, Silhavy TJ (1982) The molecular components of the signal sequence that function in the initation of protein export. J Cell Biol 95: 689–696

    Article  PubMed  CAS  Google Scholar 

  • Emr SD, Schwartz M, Silhavy TJ (1978) Mutations altering the cellular localization of the phage λ receptor, an Escherichia coli outer membrane protein. Proc Natl Acad Sci USA 75: 5802–5806

    Article  PubMed  CAS  Google Scholar 

  • Emr SD, Hanley-Way S, Silhavy TJ (1981) Suppressor mutations that restore export of a protein with a defective signal sequence. Cell 23: 79–88

    Article  PubMed  CAS  Google Scholar 

  • Ferro-Novick S, Honma M, Beckwith J (1984) The product of gene secC is involved in the synthesis of exported proteins in E. coli. Cell 38: 211–217

    Article  PubMed  CAS  Google Scholar 

  • Froshauer S (1985) Structure, localization and regulation of MalF, an inner membrane protein for maltose transport in Escherichia coli. PhD Thesis, Harvard University

    Google Scholar 

  • Froshauer S, Beckwith J (1984) The nucleotide sequence of the gene for malF protein, an inner membrane component of the maltose transport system of Escherichia coli. J Biol Chem 259: 10896–10903

    PubMed  CAS  Google Scholar 

  • Ghrayeb J, Inouye M (1984) Nine amino acid residues at the NH2-terminal of lipoprotein are sufficient for its modification, processing, and localization in the outer membrane of Escherichia coli. J Biol Chem 259: 463–467

    PubMed  CAS  Google Scholar 

  • Ghrayeb J, Kimura H, Takahara M, Hsiung H, Masui Y, Inouye M (1984) Secretion cloning vectors in Escherichia coli. EMBO J 3: 2437–2442

    PubMed  CAS  Google Scholar 

  • Gray GL, McKeown KA, Jones JSJ, Seeburg PH, Heyneker HL (1984) Pseudomonas aeruginosa secretes and correctly processes human growth hormone. Biotechnology Feb: 161–165

    Google Scholar 

  • Guarente L (1984) Use of gene fusions to study biological problems. In: Setlow J, Hollaender A (eds) Genetic engineering. Plenum, New York 6: 233–251

    Google Scholar 

  • Hall MN, Schwartz M, Silhavy TJ (1982) Sequence information within the lamB gene is required for proper routing of the bacteriophage λ receptor protein to the outer membrane of Escherichia coli K-12. J Mol Biol 156: 93–112

    Article  PubMed  CAS  Google Scholar 

  • Hayashi S, Wu HC (1983) Biosynthesis of Bacillus licheniformis penicillinase in Escherichia coli and Bacillus subtilis. J Bacteriol 156: 773–777

    PubMed  CAS  Google Scholar 

  • Hoffman C, Wright A (1985) Fusions of secreted proteins to alkaline phosphatase: an approach for studying protein secretion. Proc Natl Acad Sci USA 82: 5107–5111

    Article  PubMed  CAS  Google Scholar 

  • Ito K (1984) Identification of the sec Y (prlA) gene product involved in protein export in Escherichia coli. MGG 197: 204–208

    PubMed  CAS  Google Scholar 

  • Ito K (1986) Genetic control of protein secretion and localization. Adv Biophys 21

    Google Scholar 

  • Ito K, Bassford PJ Jr, Beckwith J (1981) Protein localization in E. coli. Is there a common step in the secretion of periplasmic and outer membrane proteins? Cell 24: 707–714

    Article  PubMed  CAS  Google Scholar 

  • Ito K, Cerretti DP, Nashimoto H, Nomura M (1984) Characterization of an amber mutation in the structural gene for ribosomal protein L5, which impairs the expression of the protein export gene, sec Y, in Escherichia coli. EMBO J 3: 2319–2324

    Google Scholar 

  • Ito K, Wittekind M, Nomura M, Shiba K, Yura T, Miura A, Nashimoto H (1983) A temperature-sensitive mutant of E. coli exhibiting slow processing of exported proteins. Cell 32: 789–797

    Article  PubMed  CAS  Google Scholar 

  • Jarvik J, Botstein D (1975) Conditional-lethal mutations that suppress genetic defects in morphogenesis by altering structural proteins. Proc Natl Acad Sci USA 72: 2738–2742

    Article  PubMed  CAS  Google Scholar 

  • Kadonaga JT, Gautier AE, Straus DR, Charles AD, Edge MD, Knowles JR (1984) The role of the β-lactamase signal sequence in the secretion of proteins by Escherichia coli. J Biol Chem 259: 2149–2154

    PubMed  CAS  Google Scholar 

  • Kiino DR, Silhavy TJ (1984) A mutation, prlF1, that relieves the lethality associated with export of β-galactosidase hybrid proteins in Escherichia coli. J Bacteriol 158: 878–883

    PubMed  CAS  Google Scholar 

  • Koshland D, Botstein D (1982) Evidence for posttranslational translocation of β-lactamase across the bacterial inner membrane. Cell 30: 893–902

    Article  PubMed  CAS  Google Scholar 

  • Koshland D, Sauer RT, Botstein D (1982) Diverse effects of mutations in the signal sequence on the secretion of β-lactamase in Salmonella typhimurium. Cell 30: 903–914

    Article  PubMed  CAS  Google Scholar 

  • Krieg P, Strachan R, Wallis E, Tabe L, Coiman A (1984) Efficient expression of cloned complementary DNAs for secretory proteins after injection into Xenopus oocytes. J Mol Biol 180: 615–643

    Article  PubMed  CAS  Google Scholar 

  • Kumamoto CA, Beckwith J (1983) Mutations in a new gene, secB, cause defective protein localization in Escherichia coli. J Bacteriol 154: 253–260

    PubMed  CAS  Google Scholar 

  • Kumamoto C, Beckwith J (1985) Evidence for specificity at an early step in protein export in Escherichia coli. J Bacteriol 163: 267–274

    PubMed  CAS  Google Scholar 

  • Kumamoto C, Oliver DB, Beckwith J (1984) Signal sequence mutations disrupt feedback between secretion of an exported protein and its synthesis in E. coli. Nature 308: 863–864

    Article  PubMed  CAS  Google Scholar 

  • Lai JS, Sarvas M, Brammer WJ, Neugebauer K, Wu HC (1981) Bacillus licheniformis penicillinase synthesized in Escherichia coli contains covalently linked fatty acid and glyceride. Proc Natl Acad Sci USA 78: 3506–3510

    Article  PubMed  CAS  Google Scholar 

  • Lee CA, Fournier MJ, Beckwith J (1985) Escherichia coli 6S RNA is not essential for growth or protein secretion. J Bacteriol 161: 1156–1161

    Google Scholar 

  • Lingappa VR, Chaidez J, Yost CS, Hedgpeth J (1984) Determinants for protein localization: β- lactamase signal sequence directs globin across microsomal membrane. Proc Natl Acad Sci USA 81: 456–460

    Article  PubMed  CAS  Google Scholar 

  • Liss LR, Oliver DB (1986) Effects of sec A mutations on the synthesis and secretion of proteins in Escherichia coli: evidence for a major export system for cell envelope proteins. J Biol Chem 261: 2299–2303

    PubMed  CAS  Google Scholar 

  • Manoil C, Beckwith J (1985) TnPhoA: a transposon probe for protein export signals. Proc Natl Acad Sci USA 82: 8129–8133

    Article  PubMed  CAS  Google Scholar 

  • Matteucci M, Lipetsky H (1986) Biotechnology 4: 51–55

    Article  CAS  Google Scholar 

  • Michaelis S, Beckwith J (1982) Mechanism of incorporation of cell envelope proteins in Escherichia coli. Ann Rev Microbiol 36: 435–465

    Article  CAS  Google Scholar 

  • Michaelis S, Guarente L, Beckwith J (1983 a) In vitro construction and characterization of phoA-lacZ gene fusions in Escherichia coli. J Bacteriol 154:356–365

    PubMed  CAS  Google Scholar 

  • Michaelis S, Inouye H, Oliver D, Beckwith J (1983 b) Mutations that alter the signal sequence of alkaline phosphatase of Escherichia coli. J Bacteriol 154: 366–374

    Google Scholar 

  • Moreno F, Fowler AV, Hall M, Silhavy TJ, Zabin I, Schwartz M (1980) A signal peptide is not sufficient to lead β-galactosidase out of the cytoplasm. Nature 286: 356–359

    Article  PubMed  CAS  Google Scholar 

  • Müller M, Blobel G (1984) Protein export in Escherichia coli requires a soluble activity. Proc Natl Acad Sci USA 81: 7737–7741

    Article  PubMed  Google Scholar 

  • Ohsuye K, Nomura M, Tanaka S, Kubota I, Nakazato H, Shinagawa H, Nakata A, Noguchi T (1983) Expression of chemically synthesized a-neo-endorphin gene fused to E. coli alkaline phosphatase. Nucleic Acids Res 11: 1283–1294

    Article  PubMed  CAS  Google Scholar 

  • Oliver DB (1985) Identification of five new essential genes involved in the synthesis of a secreted protein in Escherichia coli. J Bacteriol 161: 285–291

    PubMed  CAS  Google Scholar 

  • Oliver D, Beckwith J (1981) E. coli mutant pleiotropically defective in the export of secreted proteins. Cell 25: 765–772

    Article  PubMed  CAS  Google Scholar 

  • Oliver D, Beckwith J (1982 a) Regulation of a membrane component required for protein secretion in Escherichia coli. Cell 30: 311–319

    Article  PubMed  CAS  Google Scholar 

  • Oliver D, Beckwith J (1982 b) Identification of a new gene (sec A) and gene product involved in the secretion of envelope proteins in Escherichia coli. J Bacteriol 150: 686–691

    PubMed  CAS  Google Scholar 

  • Palva I, Sarvas M, Lehtovaara P, Shakow M, Kaariainen L (1982) Secretion of Escherichia coli β-lactamase from Bacillus subtilis by the aid of α-amylase signal sequence. Proc Natl Acad Sci USA 79: 5582–5586

    Article  PubMed  CAS  Google Scholar 

  • Palva I, Lehtovaara P, Kaariainen L, Sibakov M, Cantell K, Schein CH, Kashiwagi K, Weissman C (1983) Secretion of interferon by Bacillus subtilis. Gene 22: 229–235

    Article  PubMed  CAS  Google Scholar 

  • Rasmussen BA, Bassford PJ Jr (1985) Both linked and unlinked mutations can alter the intracellular site of synthesis of exported proteins of Escherichia coli. J Bacteriol 161: 258–264

    PubMed  CAS  Google Scholar 

  • Rasmussen BA, Bankaitis VA, Bassford PJ Jr (1984) Export and processing of malE-lacZ hybrid proteins in Escherichia coli. J Bacteriol 160: 612–617

    PubMed  CAS  Google Scholar 

  • Sarvas MO, Palva IA (1983) The penicillinase of Bacillus licheniformis is an outer membrane protein in Escherichia coli. J Bacteriol 155: 657–663

    PubMed  CAS  Google Scholar 

  • Schultz J, Silhavy TJ, Berman ML, Fiil N, Emr SC (1982) A previously unidentified gene in the spc operon of Escherichia coli K12 specifies a component of the protein export machinery. Cell 31: 227–235

    Article  Google Scholar 

  • Schwartz M (1985) Gene fusions in bacteria. In: Scaife J, Galizzi A Genetics of bacteria. Academic, London, pp 65–84

    Google Scholar 

  • Shiba K, Ito K, Yura T (1984 a) Mutation that suppresses the protein export defect of the secY mutation and causes cold-sensitive growth of Escherichia coli. J Bacteriol 160: 696–701

    PubMed  CAS  Google Scholar 

  • Shiba K, Ito K, Yura T, Cerretti DP (1984 b) A defined mutation in the protein export gene within the spc ribosomal protein operon of Escherichia coli: isolation and characterization of a new temperature-sensitive sec Y mutant. EMBO J 3: 631–635

    PubMed  CAS  Google Scholar 

  • Shiroza T, Nakazawa K, Tashiro K, Yamane K, Yanagi K, Yamasaki M, Tamura G, Saito H, Karvade Y, Taniguchi T (1985) Synthesis and secretion of active mouse interferon-B using a Bacillus subtilis α-amylase secretion vector. Gene 34: 1–8

    Article  PubMed  CAS  Google Scholar 

  • Silhavy TJ, Beckwith JR (1985) Uses of lac fusions for the study of biological problems. Microbiol Rev 43: 398–418

    Google Scholar 

  • Silhavy TJ, Casadaban MJ, Shuman HA, Beckwith JR (1976) Conversion of β-galactosidase to a membrane-bound state by gene fusion. Proc Natl Acad Sci USA 73: 3423–3427

    Article  PubMed  CAS  Google Scholar 

  • Silhavy TJ, Shuman HA, Beckwith J, Schwartz M (1977) Use of gene fusions to study outer membrane protein localization in Escherichia coli. Proc Natl Acad Sci USA 74: 5411–5415

    Article  PubMed  CAS  Google Scholar 

  • Silhavy TJ, Bassford PJ, Beckwith J (1979) A genetic approach to the study of protein localization in E. coli. In: Inouye M (ed) Bacterial outer membrane: biosynthesis, assembly and functions. Wiley, New York, pp 203–254

    Google Scholar 

  • Sodergren EJ, Davidson J, Taylor RK, Silhavy TJ (1985) Selection for mutants altered in the expression or export of outer membrane porin OmpF. J Bacteriol 162: 1047–1053

    PubMed  CAS  Google Scholar 

  • Tabe L, Krieg P, Strachan R, Jackson D, Wallis E, Coiman A (1984) Segregation of mutant ovalbumins and ovalbumin-globin fusion proteins in Xenopus oocytes. J Mol Biol 180: 645–66

    Article  PubMed  CAS  Google Scholar 

  • Takahara M, Hibler DW, Barr PJ, Inouye M (1985) The ompA signal peptide directed secretion of staphyloccoccal nuclease A by Escherichia coli. J Biol Chem 260: 2670–2674

    PubMed  CAS  Google Scholar 

  • Talmadge K, Kaufman J, Gilbert W (1980 a) Bacteria mature preproinsulin to proinsulin. Proc Natl Acad Sci USA 77: 3988–3992

    Google Scholar 

  • Talmadge K, Stahl S, Gilbert W (1980 b) Eukaryotic signal sequence transports insulin antigen in Escherichia coli. Proc Natl Acad Sci USA 77: 3369–3373

    Google Scholar 

  • Talmadge K, Brosius J, Gilbert W (1981) An internal signal sequence directs secretion and processing of proinsulin in bacteria. Nature 294: 176–178

    Article  PubMed  CAS  Google Scholar 

  • Tommassen J, van Tol H, Lugtenberg B (1983) The ultimate locatization of an OM protein of Escherichia coli Kl2 is not determined by the signal sequence. EMBO J 2: 1275–1279

    PubMed  CAS  Google Scholar 

  • Tommassen J, Pugsley AP, Korteland J, Verbakel J, Lugtenberg B (1984) Gene encoding a hybrid OmpF-PhoE pore protein in the outer membrane of Escherichia coli K12. MGG 197: 503–508

    PubMed  CAS  Google Scholar 

  • Tommassen J, Leunissen J, van Damme-Jongsten M, Overduin P (1985) Failure of E. coli K-12 to transport PhoE-LacZ hybrid protein out of the cytoplasm. EMBO J 4: 1041–1047

    PubMed  CAS  Google Scholar 

  • Ulmanen I, Lundstron K, Lehtovaara P, Sarvas M, Ruohonen M, Palva I (1985) Transcription and translation of foreign genes in Bacillus subtilis by the aid of a secretion vector. J Bacteriol 162: 176–182

    PubMed  CAS  Google Scholar 

  • von Heijne G (1980) Trans-membrane translocation of proteins: a detailed physico-chemical analysis. Eur J Biochem 103: 431–438

    Article  Google Scholar 

  • von Heijne G (1984) Analysis of the distribution of charged residues in the N-terminal region of signal sequences: implications for protein export in prokaryotic and eukaryotic cells. EMBO J 3: 2315–2318

    Google Scholar 

  • Walter P, Blobel G (1980) Purification of a membrane-associated protein complex required for protein translocation across the endoplasmic reticulum. Proc Natl Acad Sci USA 77: 7112–7116

    Article  PubMed  CAS  Google Scholar 

  • Walter P, Blobel G (1982) Signal recognition particle contains a 7S RNA essential for protein translocation across the endoplasmic reticulum. Nature 299: 691–698

    Article  PubMed  CAS  Google Scholar 

  • Walter P, Blobel G (1983) Disassembly and reconstitution of signal recognition particle. Cell 34: 525–533

    Article  PubMed  CAS  Google Scholar 

  • Zemel-Dreasen O, Zamir A (1984) Secretion and processing of an immunoglobulin light chain in Escherichia coli. Gene 27: 315–322

    Article  PubMed  CAS  Google Scholar 

Download references

Author information

Author notes

  1. S. Ferro-Novick

    Present address: Department of Cell Biology, Yale University, School of Medicine, New Haven, CT, 06510, USA

Authors and Affiliations

  1. Department of Microbiology and Molecular Genetics, Harvard Medical School, Boston, MA, 02115, USA

    J. Beckwith

Authors
  1. J. Beckwith
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. S. Ferro-Novick
    View author publications

    You can also search for this author in PubMed Google Scholar

Editor information

Editors and Affiliations

  1. Uniformed Services University of the Health Sciences, 4301 Jones Bridge Road, Bethesda, MD, 20814-4799, USA

    Henry C. Wu M.D. Ph.D (Professor of Microbiology) (Professor of Microbiology)

  2. Department of Metabolic Regulation, Boston Biomedical Research Institute, 20 Staniford Street, Boston, MA, 02114, USA

    Phang C. Tai Ph.D (Senior Staff Scientist, Associate Professor) (Senior Staff Scientist, Associate Professor)

  3. Department of Microbiology and Molecular Genetics, Harvard Medical School, 23 Shattuck Street, Boston, MA, 02115, USA

    Phang C. Tai Ph.D (Senior Staff Scientist, Associate Professor) (Senior Staff Scientist, Associate Professor)

Rights and permissions

Reprints and permissions

Copyright information

© 1986 Springer-Verlag Berlin · Heidelberg

About this paper

Cite this paper

Beckwith, J., Ferro-Novick, S. (1986). Genetic Studies on Protein Export in Bacteria. In: Wu, H.C., Tai, P.C. (eds) Protein Secretion and Export in Bacteria. Current Topics in Microbiology and Immunology, vol 125. Springer, Berlin, Heidelberg. https://doi.org/10.1007/978-3-642-71251-7_2

Download citation

  • DOI: https://doi.org/10.1007/978-3-642-71251-7_2

  • Publisher Name: Springer, Berlin, Heidelberg

  • Print ISBN: 978-3-642-71253-1

  • Online ISBN: 978-3-642-71251-7

  • eBook Packages: Springer Book Archive

Publish with us

Policies and ethics

Search

Navigation