Summary
A protein of Mr 47,000 is synthesized in Escherichia coli minicells, when these harbor a multicopy plasmid carrying IS4 in either orientation and between different flanking sequences. The protein corresponds to the sequence predicted from the known DNA sequence of IS4, as shown by partial N-terminal radiolabel protein sequence analysis. Its apparent molecular weight, however, as determined from its electrophoretic mobility in SDS polyacrylamide gels, is smaller than predicted. When compared with other plasmid-encoded proteins, the IS4-encoded protein is synthesized in minicells in small amounts. Its synthesis has not been detected in a DNA-dependent cell-free system.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Besemer J, Herpers M (1977) Suppression of insertion mutations within the gal operon of E. coli. Mol Gen Genet 151:295–304
Beyreuther K, Ehring R, Overath P, Wright JK (1980a) Microsequence analysis of lactose permease of Escherichia coli. In: Methods in peptide and protein sequence analysis. Elsevier/North-Holland Biomedical Press Amsterdam, New York, Oxford, pp 199–212
Beyreuther K, Bieseler B, Ehring R, Grießer H-W, Mieschendahl M, Müller-Hill B, Triesch I (1980b) Investigation of structure and function of lactose permease of Escherichia coli, Biochem Soc Trans 8:675–676
Calos MP, Miller JH (1980) Transposable elements. Cell 20:579–595
Chou PY, Fasman GD (1978) Empirical predictions of protein conformation. Annu Rev Biochem 47:251–276
Clewell DB, Helinski DR (1969) Supercoiled circular DNA-protein complex in Escherichia coli: Purification and induced conversion to an open circular DNA form. Proc Natl Acad Sci USA 62:1159–1166
Cohen SN, Chang ACY, Hsu L (1972) Nonchromosomal antibiotic resistance in bacteria: Genetic transformation of Escherichia coli by R-factor DNA. Proc Natl Acad Sci USA 69:2110–2114
Ehring R, Beyreuther K, Wright JK, Overath P (1980) In vitro and in vivo products of Escherichia coli lactose permease gene are identical. Nature 283:537–540
Ghosal D, Sommer H, Saedler H (1979) Nucleotide sequence of the transposable DNA element IS2. Nucl Acid Res 6:1111–1122
Grindley DF, Joyce CM (1980) Genetic and DNA sequence analysis of the kanamycin resistance transposon Tn 903. Proc Natl Acad Sci USA 77:7176–7180
Habermann P, Klaer R, Kühn S, Starlinger P, (1979) IS4 is found between eleven or twelve base pair duplications. Mol Gen Genet 175:369–373
Klaer R, Kühn S, Fritz H-J, Tillmann E, Saint-Girons I, Habermann P, Pfeifer D, Starlinger P (1980) Studies on IS4. Cold Spring Harbor Symp Quant Biol 45: in press
Klaer R, Kühn S, Tillmann E, Fritz H-J, Starlinger P (1981) The sequence of IS4. Molec Gen Genet in press
Klaer R, Starlinger P (1980) IS4 at its chromosomal site in Escherichia coli K12. Mol Gen Genet 178:285–291
Kleckner N, Ross DG (1980) recA-dependent genetic switch generated by transposon Tn10. J Mol Biol 144:215–221
Koshland D, Botstein D (1980) Secretion of beta-lactamase requires the carboxy end of the protein. Cell 20:749–760
Kühn S, Fritz H-J, Starlinger P (1979) Close vicinity of IS1 integration sites in the leader sequence of the gal operon of Escherichia coli. Mol Gen Genet 167:235–241
Lugtenberg B, Meijers J, Peters R, van der Hoek P, van Alphen L (1975) Electrophoretic resolution of the major outer membrane protein of Escherichia coli K12 into four bands. FEBS Lett 58:254–258
Mandel M, Higa A (1970) Calcium-dependent bacteriophage DNA infection. J Mol Biol 53:159–162
Mertens G, Reeve JN (1977) Synthesis of cell envelope components by anucleate cells (minicells) of Bacillus subtilis. J Bact 129:1198–1207
Miller JH (1972) Experiments in molecular genetics. Cold Spring Harbor Laboratory, New York
Ohtsubo H, Ohtsubo E (1978) Nucleotide sequence of an insertion element, IS1. Proc Natl Acad Sci USA 75:615–619
Reeve JN (1977) Bacteriophage infection of minicells. A general method for identification of “in vivo” bacteriophage directed polypeptide biosynthesis. Mol Gen Genet 158:73–79
Rothstein SJ, Jorgensen RA, Postle K, Reznikoff WS (1980) The inverted repeats of Tn5 are functionally different. Cell 19:795–805
Sharp PA, Cohen SN, Davidson N (1973) Electron microscope heteroduplex studies of sequence relations among plasmids of Escherichia coli. II. Structure of drug resistance (R) factors and F factors. J Mol Biol 75:235–255
Starlinger P (1980) IS elements and transposons. Plasmid 3:241–259
Sutcliffe JG (1978) pBR322 restriction map derived from the DNA sequence: accurate DNA size markers up to 4361 nucleotide pairs long. Nucl Acid Res 5:2721–2728
Wetekam W, Staack K, Ehring R (1971) DNA-dependent in vitro synthesis of enzymes of the galactose operon of Escherichia coli. Mol Gen Genet 112:14–27
Wetekam W, Staack K, Ehring R (1972) Relief of polarity in DNA-dependent cell-free synthesis of enzymes of the galactose operon of Escherichia coli. Mol Gen Genet 116:258–276
Zubay G, Chambers DA, Cheong LC (1970) Cell-free studies on the regulation of the lac operon. In: Beckwith JR, Zipser D (eds) The lactose operon. Cold Spring Harbor Laboratory, New York, pp 375–391
Author information
Authors and Affiliations
Additional information
Communicated by E. Bautz
Rights and permissions
About this article
Cite this article
Trinks, K., Habermann, P., Beyreuther, K. et al. An IS4-encoded protein is synthesized in minicells. Molec. Gen. Genet. 182, 183–188 (1981). https://doi.org/10.1007/BF00269656
Received:
Issue Date:
DOI: https://doi.org/10.1007/BF00269656