Summary
Insertion elements IS1 and IS2 integrated within thegal operator-promoter region, an IS1 element in genegalT and insertions IS1 and IS2 integrated in thexycIIOP region of phage λ were transcribed in vitro withE. coli RNA-polymerase. The insertion elements are transcribed exclusively by polymerase molecules started at thegal promoter and the λPR promoter respectively. No promoter exists on IS1 or IS2 which can be recognized by RNA-polymerase in the pure in vitro transcription system used.
Both insertions apparently are transcribed with a lower elongation rate thangal operon DNA or λDNA. RNAs transcribed from the termini of IS1 and IS2 respectively were analysed by hybridization experiments. They are different in sequence.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Adhya, S.L., Gottesman, M., Crombrugghe, B.de, Court, D.: Control of transcription termination. In: RNA polymerase (eds. M. Chamberlin, R. Losick, New York: Cold Spring Harbor Laboratory 1976
Anderson, W.B., Schneider, A.B., Emmer, M., Perlman, R.L., Pastan, I.: Purification of and properties of the cyclic adenosine 3′5′-monophosphate receptor protein which mediates cyclic adenosine 3′5′-monophosphate-dependent gene transcription inEscherichia coli. J. biol. Chem.246, 5929 (1971)
Besemer, J.: Suppression of gal operator-promoter insertion mutations. In: DNA insertions, plasmids and episomes (eds. A. Bukhari, J. Shapiro, S. Adhya). New York: Cold Spring Harbor Laboratory 1976
Besemer, J., Herpers, M.: Suppression of polarity of insertion mutations within thegal operon ofE. coli, Molec. gen. Genet., in press
Besemer, J., Kubai, D.F.: Isolation and characterization of ϕ80dgal transducing phages that carrygal operator-promoter insertion mutations. Molec. gen. Genet.148, 79 (1976)
Besemer, J., Wallenstein, H.-W.: Insertion IS1 can change the structure of a promoter without impairing its functionin vitro. 10. Int. Congress Biochemistry, Hamburg 1976, book of abstracts, and manuscript in prep.
Blattner, F.R., Dahlberg, J.E.: RNA synthesis starpoints in bacteriophage λ: are promoter and operator transcribed? Nature (Lond.) New Biol.237, 227 (1972)
Bøvre, K., Szybalski, W.: Multistep DNA-RNA hybridization techniques. In: Methods in enzymology, vol. 21, p. 350 (eds. L. Grosman, K. Moldave). New York and London: Academic Press 1971
Brachet, P., Eisen, H., Rambach, A.: Mutations of coliphage λ affecting the expression of replicative functionsO andP. Molec. gen. Genet.108, 266 (1970)
Bukhari, A.J., Taylor, A.L.: Influence of insertions on packaging of host sequences covalently linked to bacteriophage Mu DNA. Proc. nat. Acad. Sci. (Wash.)72, 4399 (1975)
Feiss, M., Adhya, S., Court, D.L.: Isolation of plaque-forming galactose-transducing strains of phagelambda. Genetics71, 189 (1972)
Fiandt, M., Hradecna, Z., Loceron, H.A., Szybalski, W.: Electron micrographic mapping of deletions, insertions, inversions and homologies in the DNAs of coliphage λ and ϕ80. In: The bacteriophage lambda (ed. A.D. Hershey). New York: Cold Spring Harbor Laboratory 1971
Hirsch, H.-J.: Elektronenmikroskopische Untersuchungen der DNA des Galaktose-operons vonEscherichia coli. Inaugural-Dissertation, Universität Köln 1973
Hirsch, H.-J., Saedler, H., Starlinger, P.: Insertion mutations in the control region of the galactose operon ofE. coli. II. Physical characterization of the mutations. Molec. gen. Genet.115, 266 (1972a)
Hirsch, H.-J., Starlinger, P., Brachet, P.: Two kinds of insertions in bacterial genes. Molec. gen. Genet.119, 191 (1972b)
Hübinger, A.: Isolierung und Charakterisierung von Deletionen im Galaktose-Operon des Phagen λpgal. Diplomarbeit, Univ. Köln 1973
Michaelis, G., Saedler, H., Venkov, P., Starlinger, P.: Two insetions in the galactose operon having different sizes but homologous DNA sequences. Molec. gen. Genet.104, 371 (1969)
Miller, J.H.: Experiments in molecular genetics. New York: Cold Spring Harbor Laboratory 1972
Miller, Z., Varmus, H.E., Parks, J.S., Perlman, R.L., Pastan, I.: Regulation of gal messenger ribonucleic acid synthesis inEscherichia coli by 3′,5′-cyclic adenosine monophosphate. J. biol. Chem.246, 2898 (1971)
Musso, R.E., Crombrugghe, B.de, Pastan, I., Skalar, J., Yot, P., Weissman, S.: The 5′-terminal nucleotide sequence of galactose messenger ribonucleic acid ofEscherichia coli. Proc. nat. Acad. Sci. (Wash.)71, 4940 (1974)
Nissley, S.P., Anderson, W.B., Gottesman, M.E., Perlman, R.L., Pastan, I.: In vitro transcription of thegal operon require cyclic adenosine monophosphate and cyclic adenosine monophosphate receptor protein. J. biol. Chem.246, 4671 (1971)
Pfeifer, D., Hirsch, H.-J., Bergmann, E., Hamlaoui, M.: Nonrandom distribution of deletion endpoints in thegal operon ofE. coli. Molec. gen. Genet.132, 203 (1974)
Pfeifer, D., Kubai Maroni, D., Habermann, P.: Specific sites for integration of IS-elements within the transferase gene of the galactose operon ofE. coli K12. In: DNA insertions, plasmids and episomes (eds. A. Bukhari, J. Shapiro, S. Adhya). New York: Cold Spring Harbor Laboratory 1976
Rak, B.:Gal mRNA initiated within IS2. I. Hybridization studies. Molec. gen. Genet.149, 135 (1976)
Saedler, H., Besemer, J., Kemper, B., Rosenwirth, B., Starlinger, P.: Insertion mutations in the control region of thegal operon ofE. coli. I. Biological characterization of the mutations. Molec. gen. Genet.115, 258 (1972)
Saedler, H., Reif, H.-J., Hu, S., Davidson, N.: IS2, a genetic element for turn-off and turn-on of gene activity inE. coli. Molec. gen. Genet.132, 265 (1974)
Saedler, H., Starlinger, P.: Induction of galactokinase by an 0°-mutant of the galactose operon inEscherichia coli. Biochem. biophys. Res. Commun.20, 647 (1965)
Saedler, H., Starlinger, P.: 0°-mutations in the galactose operon ofE. coli. I. Genetic characterization. Molec. gen. Genet.100, 178 (1967a)
Saedler, H., Starlinger, P.: 0°-mutations in the galactose operon ofE. coli. II. Physiological characterization. Molec. gen. Genet.100, 190 (1967b)
Schmidt, F., Besemer, J., Starlinger, P.: The isolation of IS1 and IS2 DNA. Molec. gen. Genet.145, 145 (1976)
Starlinger, P., Saedler, H.: IS-elements in microorganisms. Current topics in microbiology and immunology, vol. 75, p. 111. Berlin-Heidelberg-New York: Springer 1976
Starlinger, P., Saedler, H., Rak, B., Tillmann, E., Venkov, P., Waltschewa, L.: mRNA distal to polar nonsense and insertion mutations in thegal operon ofE. coli. Molec. gen. Genet.122, 279 (1973)
Szybalski, W., Kubinski, H., Hradecna, Z., Summers, W.: Analytical and preparative separation of the complementary DNA strands. In: Methods in enzymology, vol. 21, p. 386 (eds. L. Grosman, K. Moldave). New York and London: Academic Press 1971
Wetekam, W., Staack, K., Ehring, R.: Relief of polarity in DNA-dependent cell-free synthesis of enzymes of the galactose operon ofEscherichia coli. Molec. gen. Genet.116, 258 (1972)
Author information
Authors and Affiliations
Additional information
Communicated by E. Bautz
Rights and permissions
About this article
Cite this article
Besemer, J., Molzberger, C. Transcription of insertion elements IS1 and IS2 in vitro. Molec. Gen. Genet. 153, 51–60 (1977). https://doi.org/10.1007/BF01035996
Received:
Issue Date:
DOI: https://doi.org/10.1007/BF01035996