In vivo and in vitro synthesis of Escherichia coli maltose-binding protein under regulatory control of the lacUV5 promoter-operator
10.1128/jb.164.2.665-673.1985
Crossref journal-article
American Society for Microbiology
Journal of Bacteriology (235)
Abstract

It has not been possible to obtain in vitro expression of the positively regulated malE gene encoding the periplasmic maltose-binding protein (MBP) of Escherichia coli. To facilitate in vitro malE expression, we constructed plasmids that place the malE gene under transcriptional control of the lacUV5 promoter-operator. These plasmids could be grouped into three classes, based upon their ability to complement in vivo a chromosomal malE deletion in the presence or absence of isopropyl thiogalactoside. In the one class I plasmid analyzed, the lacUV5-malE junction was just 3' to the malE ATG initiation codon, and this plasmid did not complement the malE deletion. Class II and class III plasmids retained various amounts of the malE promoter. MBP synthesis was solely under control of the lacUV5 promoter in the class II plasmids, and MBP synthesis was under control of both the lacUV5 and malE promoters in the class III plasmids. A malE mutation that renders the MBP signal peptide export defective was genetically recombined onto one of the class II plasmids. The in vivo synthesis and export of plasmid-encoded MBP were studied in the presence and absence of isopropyl thiogalactoside and maltose and in a strain harboring a prlA mutation that suppresses the malE signal sequence mutation and is thought to alter the export machinery of cells. In addition, both class II and class III plasmids programmed the synthesis of precursor MBP in an in vitro-coupled transcription-translation system. When precursor MBP was synthesized in vitro in the presence of E. coli membrane vesicles, a significant portion of wild-type precursor MBP, but not export-defective precursor MBP, was converted to a form that migrated on sodium dodecyl sulfate-polyacrylamide gels identically to mature MBP synthesized in vivo.

Bibliography

Rasmussen, B. A., MacGregor, C. H., Ray, P. H., & Bassford, P. J. (1985). In vivo and in vitro synthesis of Escherichia coli maltose-binding protein under regulatory control of the lacUV5 promoter-operator. Journal of Bacteriology, 164(2), 665–673.

Authors 4
  1. B A Rasmussen (first)
  2. C H MacGregor (additional)
  3. P H Ray (additional)
  4. P J Bassford (additional)
References 34 Referenced 33
  1. Bankaitis , V. A. , and P. J. Bassford , Jr. 1985 . Proper interaction between at least two components is required for efficient export of proteins to the Escherichia coli cell envelope . J. Bacteriol. 161 : 169 - 178 . (10.1128/JB.161.1.169-178.1985) / J. Bacteriol. / Proper interaction between at least two components is required for efficient export of proteins to the Escherichia coli cell envelope by Bankaitis V. A. (1985)
  2. Bankaitis , V. A. , B. A. Rasmussen , and P. J. Bassford , Jr. 1984 . Intragenic suppressor mutations that restore export of maltose binding protein with a truncated signal peptide , Cell 37 : 243 - 252 . (10.1016/0092-8674(84)90320-9) / Cell / Intragenic suppressor mutations that restore export of maltose binding protein with a truncated signal peptide by Bankaitis V. A. (1984)
  3. Bankaitis , V. A. , J. P. Ryan , B. A. Rasmussen , and P. J. Bassford , Jr. 1985 . The use of genetic techniques to analyze protein export in Escherichia coli . Curr. Top. Membr. Transp. 24 : 105 - 150 . (10.1016/S0070-2161(08)60325-1) / Curr. Top. Membr. Transp. / The use of genetic techniques to analyze protein export in Escherichia coli by Bankaitis V. A. (1985)
  4. Bassford , P. , and J. Beckwith . 1979 . Escherichia coli mutants accumulating the precursor of a secreted protein in the cytoplasm . Nature (London) 277 : 538 - 541 . (10.1038/277538a0) / Nature (London) / Escherichia coli mutants accumulating the precursor of a secreted protein in the cytoplasm by Bassford P. (1979)
  5. Bedouelle , H. 1983 . Mutations in the promoter of the malEFG and malK-lamB operons of Escherichia coli K12 . J. Mol. Biol. 170 : 861 - 882 . (10.1016/S0022-2836(83)80192-2) / J. Mol. Biol. / Mutations in the promoter of the malEFG and malK-lamB operons of Escherichia coli K12 by Bedouelle H. (1983)
  6. Bedouelle , H. , P. J. Bassford , Jr. , A. V. Fowler , I. Zabin , J. Beckwith , and M. Hofnung . 1980 . Mutations which alter the function of the signal sequence of the maltose binding protein of Escherichia coli . Nature (London) 285 : 78 - 81 . (10.1038/285078a0) / Nature (London) / Mutations which alter the function of the signal sequence of the maltose binding protein of Escherichia coli by Bedouelle H. (1980)
  7. Bedouelle , H. , and M. Hofnung . 1982 . A DNA sequence containing the control regions of the malEFG and malK-lamB operons in Escherichia coli K12 . Mol. Gen. Genet. 185 : 82 - 87 . (10.1007/BF00333794) / Mol. Gen. Genet. / A DNA sequence containing the control regions of the malEFG and malK-lamB operons in Escherichia coli K12 by Bedouelle H. (1982)
  8. Casadaban , M. J. 1976 . Transposition and fusion of the lac operon to selected promoters in E. coli using bacteriophage lambda and Mu . J. Mol. Biol. 104 : 541 - 555 . (10.1016/0022-2836(76)90119-4) / J. Mol. Biol. / Transposition and fusion of the lac operon to selected promoters in E. coli using bacteriophage lambda and Mu by Casadaban M. J. (1976)
  9. Chen , L. , D. Rhoads , and P. C. Tai . 1985 . Alkaline phosphatase and OmpA protein can be translocated posttranslationally into membrane vesicles of Escherichia coli . J. Bacteriol. 161 : 973 - 980 . (10.1128/JB.161.3.973-980.1985) / J. Bacteriol. / Alkaline phosphatase and OmpA protein can be translocated posttranslationally into membrane vesicles of Escherichia coli by Chen L. (1985)
  10. Debarbouile , M. , and M. Schwartz . 1979 . The use of gene fusions to study the expression of malT, the positive regulator gene of the maltose regulon . J. Mol. Biol. 132 : 521 - 534 . (10.1016/0022-2836(79)90273-0) / J. Mol. Biol. / The use of gene fusions to study the expression of malT, the positive regulator gene of the maltose regulon by Debarbouile M. (1979)
  11. DiRienzo , J. M. , K. Nakamura , and M. Inouye . 1978 . The outer membrane proteins of gram negative bacteria: biosynthesis, assembly, and functions. Annu . Rev. Biochem. 47 : 481 - 532 . (10.1146/annurev.bi.47.070178.002405) / Rev. Biochem. / The outer membrane proteins of gram negative bacteria: biosynthesis, assembly, and functions. Annu by DiRienzo J. M. (1978)
  12. Duplay , P. , H. Bedoueile , A. Fowler , I. Zabin , W. Saurin , and M. Hofnung . 1984 . Sequences of the malE gene and of its product, the maltose-binding protein of Escherichia coli K12 . J. Biol. Cherh. 259 : 10606 - 10613 . (10.1016/S0021-9258(18)91005-7) / J. Biol. Cherh. / Sequences of the malE gene and of its product, the maltose-binding protein of Escherichia coli K12 by Duplay P. (1984)
  13. Emr , S. D. , and P. J. Bassford , 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 . (10.1016/S0021-9258(19)83857-7) / J. Biol. Chem. / Localization and processing of outer membrane and periplasmic proteins in Escherichia coli strains harboring export-specific suppressor mutations by Emr S. D. (1982)
  14. Emr , S. D. , S. Hanley-Way , and T. J. S Hihavy . 1981 . Suppressor mutations that restore export of a protein with a defective signal sequence . Cell 23 : 79 - 88 . (10.1016/0092-8674(81)90272-5) / Cell / Suppressor mutations that restore export of a protein with a defective signal sequence by Emr S. D. (1981)
  15. Ferro-Novick , S. , M. Honmd , and J. Beckwith . 1984 . The product of gene secC is involved in the synthesis of exported proteins in E. coli . Cell 38 : 211 - 217 . (10.1016/0092-8674(84)90542-7) / Cell / The product of gene secC is involved in the synthesis of exported proteins in E. coli by Ferro-Novick S. (1984)
  16. Josefsson , L. - G. , and L. L. Randall . 1981 . Processing in vivo of precursor maltose binding protein in Escherichia coli occurs post-translationally as well as co-translationally . J. Biol. Chem. 256 : 2504 - 2507 . (10.1016/S0021-9258(19)69811-X) / J. Biol. Chem. / Processing in vivo of precursor maltose binding protein in Escherichia coli occurs post-translationally as well as co-translationally by Josefsson L. (1981)
  17. Kellermann , O. , and S. Szmelcman . 1974 . Active transport of maltose in Escherichia coli K12: involvement of a "periplasmic " maltose binding protein. Eur. J. Biochem. 47 : 139 - 149 . (10.1111/j.1432-1033.1974.tb03677.x) / Eur. J. Biochem. / Active transport of maltose in Escherichia coli K12: involvement of a "periplasmic by Kellermann O. (1974)
  18. Lauer , G. , R. Dastrana , J. Sherley , and M. Ptashne . 1981 . Construction of overproducers of the bacteriophage 434 repressor and cro proteins . J. Mol. Appl. Genet. 1 : 139 - 147 . / J. Mol. Appl. Genet. / Construction of overproducers of the bacteriophage 434 repressor and cro proteins by Lauer G. (1981)
  19. Maniatis T. E. F. Fritsch and J. Sambrook. 1982. Molecular cloning. Cold Spring Harbor Laboratory Cold Spring Harbor N.Y.
  20. Marchal , C. , J. Greenblatt , and M. Hofnung . 1978 . malB region in Escherichia coli K-12: specialized transducing bacteriophages and first restriction map . J. Bacteriol. 136 : 1109 - 1119 . (10.1128/JB.136.3.1109-1119.1978) / J. Bacteriol. / malB region in Escherichia coli K-12: specialized transducing bacteriophages and first restriction map by Marchal C. (1978)
  21. Marchal , C. , D. Perrin , J. Hedgpeth , and M. Hofnung . 1980 . Synthesis and maturation of lambda receptor in Escherichia coli K-12: in vivo and in vitro expression of gene lamB under lac promoter control . Proc. Natl. Acad. Sci. USA 77 : 1491 - 1495 . (10.1073/pnas.77.3.1491) / Proc. Natl. Acad. Sci. USA / Synthesis and maturation of lambda receptor in Escherichia coli K-12: in vivo and in vitro expression of gene lamB under lac promoter control by Marchal C. (1980)
  22. Messing , J. 1983 . New M13 vectors for cloning . Methods Enzymol. 101 : 20 - 78 . (10.1016/0076-6879(83)01005-8) / Methods Enzymol. / New M13 vectors for cloning by Messing J. (1983)
  23. Miller J. H. 1972. Experiments in molecular genetics. Cold Spring Harbor Laboratory Cold Spring Harbor N.Y.
  24. Model , P. , and M. Russel . 1983 . Insertion of proteins into bacterial membranes . Methods Enzymol. 97 : 139 - 146 . / Methods Enzymol. / Insertion of proteins into bacterial membranes by Model P. (1983)
  25. Muller , M. , and G. Blobel . 1984 . In vitro translocation of bacterial proteins across the plasma membrane of Escherichia coli . Proc. Natl. Acad. Sci. USA 81 : 7421 - 7425 . (10.1073/pnas.81.23.7421) / Proc. Natl. Acad. Sci. USA / In vitro translocation of bacterial proteins across the plasma membrane of Escherichia coli by Muller M. (1984)
  26. Oliver , D. B. , and J. Beckwith . 1982 . Regulation of a membrane component required for secretion in Escherichia coli . Cell 30 : 311 - 319 . (10.1016/0092-8674(82)90037-X) / Cell / Regulation of a membrane component required for secretion in Escherichia coli by Oliver D. B. (1982)
  27. Randall , L. L. 1983 . Translocation of domains of nascent periplasmic proteins across the cytoplasmic membrane is independent of elongation . Cell 33 : 231 - 240 . (10.1016/0092-8674(83)90352-5) / Cell / Translocation of domains of nascent periplasmic proteins across the cytoplasmic membrane is independent of elongation by Randall L. L. (1983)
  28. Randall , L. L. , and S. J. S. Hardy . 1977 . Synthesis of exported proteins by membrane-bound polysomes from Escherichia coli . Eur. J. Biochem. 75 : 43 - 53 . (10.1111/j.1432-1033.1977.tb11502.x) / Eur. J. Biochem. / Synthesis of exported proteins by membrane-bound polysomes from Escherichia coli by Randall L. L. (1977)
  29. Randall L. L. and S. J. S. Hardy. 1984. Export of protein in bacteria: data and dogma p. 1-10. In B. Satir (ed.) Modem cell biology vol. 3. Alan R. Liss Inc. New York.
  30. Randall , L. L. , S. J. S. Hardy , and L. -G. Josefsson . 1978 . Precursors of three exported proteins in Escherichia coli . Proc. Natl. Acad. Sci. USA 75 : 1209 - 1212 . (10.1073/pnas.75.3.1209) / Proc. Natl. Acad. Sci. USA / Precursors of three exported proteins in Escherichia coli by Randall L. L. (1978)
  31. Rasmussen , B. A. , and P. J. Bassford , 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 . (10.1128/JB.161.1.258-264.1985) / J. Bacteriol. / Both linked and unlinked mutations can alter the intracellular site of synthesis of exported proteins of Escherichia coli by Rasmussen B. A. (1985)
  32. Sanger , F. , S. Nicklen , and A. R. Carlson . 1977 . DNA sequencing with chain terminating inhibitors . Proc. Natl. Acad. Sci. USA 74 : 5463 - 5467 . (10.1073/pnas.74.12.5463) / Proc. Natl. Acad. Sci. USA / DNA sequencing with chain terminating inhibitors by Sanger F. (1977)
  33. Silhavy , T. J. , S. A. Benson , and S. D. Emr . 1983 . Mechanisms of protein localization . Microbiol. Rev. 47 : 313 - 344 . (10.1128/mr.47.3.313-344.1983) / Microbiol. Rev. / Mechanisms of protein localization by Silhavy T. J. (1983)
  34. Sihavy T. J. M. L. Berman and L. W. Enquist. 1984. Experiments with gene fusions. Cold Spring Harbor Laboratory Cold Spring Harbor N.Y.
Dates
Type When
Created 5 years, 8 months ago (Jan. 3, 2020, 1:23 p.m.)
Deposited 4 years, 1 month ago (July 29, 2021, 1:29 p.m.)
Indexed 1 year, 7 months ago (Jan. 12, 2024, 4:37 a.m.)
Issued 39 years, 10 months ago (Nov. 1, 1985)
Published 39 years, 10 months ago (Nov. 1, 1985)
Published Print 39 years, 10 months ago (Nov. 1, 1985)
Funders 0

None

@article{Rasmussen_1985, title={In vivo and in vitro synthesis of Escherichia coli maltose-binding protein under regulatory control of the lacUV5 promoter-operator}, volume={164}, ISSN={1098-5530}, url={http://dx.doi.org/10.1128/jb.164.2.665-673.1985}, DOI={10.1128/jb.164.2.665-673.1985}, number={2}, journal={Journal of Bacteriology}, publisher={American Society for Microbiology}, author={Rasmussen, B A and MacGregor, C H and Ray, P H and Bassford, P J}, year={1985}, month=nov, pages={665–673} }