Cotranslational Secretion of Diphtheria Toxin and Alkaline Phosphatase in Vitro: Involvement of Membrane Protein(s)

Overview

Journal J Bacteriol

Publisher American Society for Microbiology

Specialty Microbiology

Date 1980 Mar 1

PMID 6988404

Citations 11

Authors

W P Smith

Affiliations

Soon will be listed here.

Abstract

Crude messenger ribonucleic acid fractions isolated from Corynebacterium diphtheriae and Escherichia coli were translated in an E. coli in vitro protein-synthesizing system and yielded precursors of the secreted proteins diphtheria toxin and alkaline phosphatase, respectively. Addition of inverted E. coli inner membrane vesicles to the system during the initial stages of translation resulted in the intravesicular segregation of mature diphtheria toxin and alkaline phosphatase. Outer membrane vesicles or inner membrane vesicles whose cytoplasmic surfaces had been treated with pronase could not mediate transmembrane transfer of diphtheria toxin or alkaline phosphatase. However, inner membrane vesicles isolated from E. coli spheroplasts which had been treated with pronase and inner membrane vesicles complexed with ribosomes during pronase treatment were functional in transmembrane transfer. At temperatures below the phase transition of E. coli membranes, no intravesicular segregation of alkaline phosphatase or diphtheria toxin was observed. The precursor forms of each protein accumulated free from the vesicles. These results suggest that an inner membrane protein, exposed on the cytoplasmic surface, plays an integral role in secretion.

Citing Articles

Regulation of toxinogenesis in Corynebacterium diphtheriae. I. Mutations in bacteriophage beta that alter the effects of iron on toxin production.

Welkos S, Holmes R J Virol. 1981; 37(3):936-45.

PMID: 6785449 PMC: 171091. DOI: 10.1128/JVI.37.3.936-945.1981.

A 64-kilodalton membrane protein of Bacillus subtilis covered by secreting ribosomes.

Horiuchi S, Tai P, Davis B Proc Natl Acad Sci U S A. 1983; 80(11):3287-91.

PMID: 6407010 PMC: 394026. DOI: 10.1073/pnas.80.11.3287.

In vitro translocation of bacterial proteins across the plasma membrane of Escherichia coli.

Muller M, Blobel G Proc Natl Acad Sci U S A. 1984; 81(23):7421-5.

PMID: 6390437 PMC: 392158. DOI: 10.1073/pnas.81.23.7421.

Mechanisms of protein localization.

Silhavy T, Benson S, Emr S Microbiol Rev. 1983; 47(3):313-44.

PMID: 6355805 PMC: 281579. DOI: 10.1128/mr.47.3.313-344.1983.

Molecular cloning and expression of gene fragments from corynebacteriophage beta encoding enzymatically active peptides of diphtheria toxin.

Tweten R, Collier R J Bacteriol. 1983; 156(2):680-5.

PMID: 6313613 PMC: 217883. DOI: 10.1128/jb.156.2.680-685.1983.

References

Palmiter R . Magnesium precipitation of ribonucleoprotein complexes. Expedient techniques for the isolation of undergraded polysomes and messenger ribonucleic acid. Biochemistry. 1974; 13(17):3606-15. DOI: 10.1021/bi00714a032. View

Osborn M, Munson R . Separation of the inner (cytoplasmic) and outer membranes of Gram-negative bacteria. Methods Enzymol. 1974; 31:642-53. DOI: 10.1016/0076-6879(74)31070-1. View

Kaback H . Transport studies in bacterial membrane vesicles. Science. 1974; 186(4167):882-92. DOI: 10.1126/science.186.4167.882. View

Glenn A . Production of extracellular proteins by bacteria. Annu Rev Microbiol. 1976; 30:41-62. DOI: 10.1146/annurev.mi.30.100176.000353. View

Inouye H, Beckwith J . Synthesis and processing of an Escherichia coli alkaline phosphatase precursor in vitro. Proc Natl Acad Sci U S A. 1977; 74(4):1440-4. PMC: 430790. DOI: 10.1073/pnas.74.4.1440. View

Randall L, Hardy S . Synthesis of exported proteins by membrane-bound polysomes from Escherichia coli. Eur J Biochem. 1977; 75(1):43-53. DOI: 10.1111/j.1432-1033.1977.tb11502.x. View

Tribhuwan R, Pradhan D . Induction of alkaline phosphatase in Escherichia coli: effect of procaine hydrochloride. J Bacteriol. 1977; 131(2):431-7. PMC: 235448. DOI: 10.1128/jb.131.2.431-437.1977. View

Smith W, Tai P, Thompson R, Davis B . Extracellular labeling of nascent polypeptides traversing the membrane of Escherichia coli. Proc Natl Acad Sci U S A. 1977; 74(7):2830-4. PMC: 431309. DOI: 10.1073/pnas.74.7.2830. View

Chang C, Blobel G, Model P . Detection of prokaryotic signal peptidase in an Escherichia coli membrane fraction: endoproteolytic cleavage of nascent f1 pre-coat protein. Proc Natl Acad Sci U S A. 1978; 75(1):361-5. PMC: 411248. DOI: 10.1073/pnas.75.1.361. View

10.

Smith W, Tai P, Davis B . Nascent peptide as sole attachment of polysomes to membranes in bacteria. Proc Natl Acad Sci U S A. 1978; 75(2):814-7. PMC: 411347. DOI: 10.1073/pnas.75.2.814. View

11.

Wickner W, Mandel G, Zwizinski C, Bates M, Killick T . Synthesis of phage M13 coat protein and its assembly into membranes in vitro. Proc Natl Acad Sci U S A. 1978; 75(4):1754-8. PMC: 392418. DOI: 10.1073/pnas.75.4.1754. View

12.

Kreibich G, Ulrich B, Sabatini D . Proteins of rough microsomal membranes related to ribosome binding. I. Identification of ribophorins I and II, membrane proteins characteristics of rough microsomes. J Cell Biol. 1978; 77(2):464-87. PMC: 2110038. DOI: 10.1083/jcb.77.2.464. View

13.

Randall L, Hardy S, Josefsson L . Precursors of three exported proteins in Escherichia coli. Proc Natl Acad Sci U S A. 1978; 75(3):1209-12. PMC: 411439. DOI: 10.1073/pnas.75.3.1209. View

14.

Pages J, Piovant M, Varenne S, Lazdunski C . Mechanistic aspects of the transfer of nascent periplasmic proteins across the cytoplasmic membrane in Escherichia coli. Eur J Biochem. 1978; 86(2):589-602. DOI: 10.1111/j.1432-1033.1978.tb12343.x. View

15.

Varenne S, Piovant M, Pages J, Lazdunski C . Evidence for synthesis of alkaline phosphatase on membrane-bound polysomes in Escherichia coli. Eur J Biochem. 1978; 86(2):603-6. DOI: 10.1111/j.1432-1033.1978.tb12344.x. View

16.

Murphy J, Michel J, Teng M . Evidence that the regulation of diphtheria toxin production is directed at the level of transcription. J Bacteriol. 1978; 135(2):511-6. PMC: 222410. DOI: 10.1128/jb.135.2.511-516.1978. View

17.

Smith W, Tai P, Davis B . Interaction of secreted nascent chains with surrounding membrane in Bacillus subtilis. Proc Natl Acad Sci U S A. 1978; 75(12):5922-5. PMC: 393088. DOI: 10.1073/pnas.75.12.5922. View

18.

Smith W, Tai P, Davis B . Extracellular labeling of growing secreted polypeptide chains in Bacillus subtilis with diazoiodosulfanilic acid. Biochemistry. 1979; 18(1):198-202. DOI: 10.1021/bi00568a030. View

19.

Smith W, Tai P, Murphy J, Davis B . Precursor in cotranslational secretion of diphtheria toxin. J Bacteriol. 1980; 141(1):184-9. PMC: 293559. DOI: 10.1128/jb.141.1.184-189.1980. View

20.

Davis B, MINGIOLI E . Mutants of Escherichia coli requiring methionine or vitamin B12. J Bacteriol. 1950; 60(1):17-28. PMC: 385836. DOI: 10.1128/jb.60.1.17-28.1950. View