Summary
Eight λilvC transducing phages generated from E. coli K12 secondary site lysogens have been analysed genetically and physically. Two of them carry, in addition, the rho gene and its promotor region, but not the cya gene. The ilvO603 mutation has been located between ilvG and ilvE. Electrophoretic analysis of the proteins synthesized by these phages in a system of UV irradiated cells allowed us to assign molecular weights of 55000 and 66000 daltons to the ilvC and the ilvD gene products, respectively, and to show that an ilvG-encoded polypeptide of 60000 daltons is made from an ilvO - but not from an ilvO + phage. The expression of the ilvG gene is discussed in the light of the recent finding of a promoter-attenuator region lying upstream to ilvG. Finally, we have found that one of the λilv phages does not have the classical structure of a transducing phage.
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Abrescia P, Guardiola J, Foresti M, Lamberti A, Iaccarino M (1979) Thereonine deaminase: Autogenous regulator of the ilv genes in Escherichia coli K-12. Mol Gen Genet 171:261–275
Arfin ST, Ratzkin B, Umbarger HE (1969) The metabolism of valine and isoleucine in Escherichia coli. XVII The role of induction in the derepression of acetohydroxy acid isomeroreductase. Biochem Biophys Res Commun 37:902–908
Bachmann BJ, Low KB (1980) Linkage map of Escherichia coli K-12, edition 6. Microbiol Rev 44:1–56
Baez M, Patin DW, Calhoun DH (1979) Deletion mapping of the ilvGEDAC genes of Escherichia coli K-12. Mol Gen Genet 169:289–297
Berg CM, Shaw KJ, Berg DE (1980) The ilvG gene is expressed in Escherichia coli K-12. Gene 12:165–170
Bonner WM, Laskey RA (1974) A film detection method for tritiumlabelled proteins and nucleic acids in polyacrylamide gels. Eur J Biochem 46:83–88
Calhoun DH, Rimerman RA, Hatfield GW (1973) Threonine deaminase from Escherichia coli. I. Purification and properties. J Biol Chem 248:3511–3516
Campbell A (1971) Genetic structure. In: Hershey AD (ed) The bacteriophage λ. Cold Spring Harbor Laboratory, New York, p 13
Childs GJ, Ohtsubo H, Ohtsubo E, Sonnenberg F, Freundlich M (1977) Restriction endonuclease mapping of the Escherichia coli K-12 chromosome in the vicinity of the ilv genes. J Mol Biol 117:175–193
Cohen BM, Jones EW (1976) New map location of ilvO in Escherichia coli. Genetics 83:201–225
Das A, Court, D., Adhya S (1976) Isolation and characterization of conditional lethal mutants of Escherichia coli defective in transcription termination factor rho. Proc Natl Acad Sci USA 73:1959–1963
Davis RM, Simon M, Davidson N (1971) Electron microscope heteroduplex methods for mapping regions of base sequence homology in nucleic acids. In: Grossman L and Moldave K (eds) Methods in enzymology, vol 21. Academic Press, New-York, p 413
De Felice M, Guardiola J, Malorni MC, Klopotowski T, Iaccarino M (1974) Regulation of the pool size of valine in Escherichia coli K-12. J Bacteriol 120:1058–1067
De Felice M, Squires C, Levinthal M, Guardiola, J., Lamberti A, Iaccarino M (1977) Growth inhibition of Escherichia coli K-12 by valine: a consequence of a regulatory pattern. Mol Gen Genet. 156:1–7
De Felice M, Levinthal M, Iaccarino M, Guardiola J (1979) Growth inhibition as a consequence of antagnonism between related amino acids: effects of valine in Escherichia coli K-12. Microbiol Rev 43:42–58
Enquist L, Tierneier D, Leder P, Weisberg R, Stenberg N (1976) Safer derivatives of bacteriophage λgt, λC for use in cloning of recombinant DNA molecules. Nature 259:597–598
Favre R, Wiater A, Puppo S, Iaccarino M, Noelle R, Freundlich M (1976) Expression of a valine-resistant acetolactate, synthase activity mediated by the ilvO and ilvG genes of Escherichia coli K-12. Mol Gen Genet 143:243–252
Franklin NC, Yanofsky C (1976) The N protein of λ: evidence bearing on transcription termination polarity and the alteration of Escherichia coli RNA polymerase. In Losick R and Chamberlin, M (eds) RNA polymerase. Cold Spring Harbor Laboratory, New York, p 693
Hopkins AS, Murray NE, Brammar WJ (1976) Characterization of λtrp transducing bacteriophages made in vitro. J Mol Biol 107:549–569
Iaccarino M, Guardiola J, De Felice M, Favre R (1978) Regulation of isoleucine and valine biosynthesis. In Current topics in cellular regulation 14: 29–73
Imai M, Shigesada K (1978) Studies on the altered rho factor in nitA mutants of Escherichia coli defective in transcription termination. I. Characterization and quantitative determination of rho in cell extracts. J Mol Biol 120:451–466
Inoko H, Shigesada K, Imai K (1977) Isolation and characterization of conditional-lethal rho mutants of Escherichia coli. Proc Natl Acad Sci USA 74:1162–1166
Jørgensen P, Collins J, Fiil N, Von Meyenburg K (1978) A ribosomal RNA gene, rrnC, of Escherichia coli, mapped by specialized transducing λdilv and λdrbs phages. Mol Gen Genet 163:223–228
Laemmli UK (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227:680–685
Lawther RP, Nichols B, Zurawski G, Hatfield GW (1979) The nucleotide sequence preceding and including the beginning of the ilvE gene of the ilvGEDA operon of Escherichia coli K-12. Nucl Acids Res 7:2289–2301
Lawther RP, Hatfield GW (1980) Multivalent translational control of transcription termination at attenuator of ilvGEDA operon of Escherichia coli K-12. Proc Natl Acad Sci USA 77:1862–1866
Lee-Peng FC, Hermodson MA, Kohlhaw GB (1979) Transaminase B from Escherichia coli: quanternary structure amino-terminal sequence, substrate specificity and absence of a separate, valine α-ketoglutarate activity. J Bacteriol 139:339–345
Lo Schiavo F, Favre R, Kasai T, Cascino A, Guardiola J, Caro L, Iaccarino M (1975) The Φ80λdilv phage and its use to detect messenger RNA related to isoleucine-valine metabolsim. J Mol Biol 99:353–368
Marsh NJ, Duggan DE (1972) Ordering of mutant sizes in the isoleucine-valine genes of Escherichia coli by use of merogenotes derived from F-14: a new procedure for fine-structure mapping. J Bacteriol 109:730–740
McCorkle GM, Leathers TD, Umbarger HE (1978) Physical organization of the ilvEDAC genes of Escherichia coli strain K-12. Proc Natl Acad Sci USA 75:89–93
Miller JH (1972) In: Experiments in molecular genetics. Cold Spring Harbor Laboratory, New York
Newman TC, Levinthal M (1980) A new map location for the ilvB locus of Escherichia coli K-12. Genetics 96:59–77
Nargang FE, Subrahmanyam CS, Umbarger HE (1980) Nucleotide sequence of the attenuator region of the ilvGEDA operon of Escherichia coli. Proc Natl Acad Sci USA 77:1823–1827
Oppenheim AB, Katzir N, Oppenheim A (1977) Regulation of protein synthesis in bacteriophage λ. Restoration of gene expression in λN - strains by mutation in the cro gene. Virologie 79:405–425
Patin DW, Calhoun DM (1979) Mapping of ilvO loci of Escherichia coli K-12 with bacteriophage λdilv. J Bacteriol 137:1234–1242
Ptashne M (1967) Isolation of the λ repressor. Proc Natl Acad Sci USA 57:306–313
Roberts JW (1969) Termination factor for RNA synthesis. Nature 224:1169–1174
Schrenk WJ, Weisberg RA (1975) A simple method for making new transducing lines of coliphage λ. Mol Gen Genet 137:101–107
Shimada K, Weisberg RA, Gottesman ME (1973) Prophage lambda at unusual chromosomal locations. II. Mutations induced by bacteriophage lambda in Escherichia coli K-12. J Mol Biol 80:297–314
Shimada K, Weisberg RA, Gottesman ME (1975) Prophage lambda at unusual chromosomal locations. III. The components of the secondary attachment sites. J Mol Biol 93:415–429
Silver L, Chandler M, Boy de la Tour E, Caro L (1977) Origin and direction of replication of the drug resistance plasmid R100-1 and of a resistance transfer factor derivative in synchronized cultures. J Bacteriol 131:929–942
Smith J, Smith F, Umbarger H (1979) Mutation affecting the formation of acetohydroxyacid synthase II in Escherichia coli K-12. Mol Gen Genet 169:299–314
Springer M, Graffe M, Hennecke H (1977) Specialized transducing phage for the initiation factor IF3 gene in E. coli. Proc Natl Acad Sci USA 74:3970–3974
Steitz JA (1979) RNA-RNA interaction during polypeptide chain initation. In: Chambliss G, Graven GR, Davies J, Davis K, Kahan L, Nomura M (eds) Ribosomes: Structure, function and genetics. University Park Pres, Baltimore, p 479
Subrahmanyam CS, McCorkle GM, Umbarger HE (1980a) Physical location of the ilvO determinant in Escherichia coli K-12 deoxyribonucleic acid. J Bacteriol 142:547–555
Subrahmanyam CS, Noti JD, Umbarger HE (1980b) Regulation of ilvEDA expression occurs upstream of ilvG in Escherichia coli: Additional evidence for an ilvGEDA operon. J Bacteriol 144:279–290
Sutcliffe JG (1978) pBR322 restriction map derived from the DNA sequence: accurate DNA size markers up to 4361 nucleotide pairs long. Nucl Acids Res 5:2721–2728
Szybalski EH, Szybalski W (1979) A comprehensive molecular map of bacteriophage lambda. Gene 7:217–270
Thomas M, Cameron JR, Davis RW (1974) Viable molecular hybrids of bacteriophage lambda and eucaryotic DNA. Proc Natl Acad Sci USA 71:4579–4583
Thomas M, Davis RW (1975) Studies on the cleavage map of bacteriophage lambda with EcoRI restriction endonuclease. J Mol Biol 91:315–328
Umbarger HE (1978) Amino acid biosynthesis and its regulation. Ann Rev Biochem 47:533–606
Watson MD, Wild J, Umbarger HE (1979) Positive control of ilvC expression in Escherichia coli K-12; identification and mapping of the ilvY regulatory gene. J. Bacteriol 139:1014–1020
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Uzan, M., Favre, R., Gallay, E. et al. Genetical and structural analysis of a group of λilv and λrho transducing phages. Molec. Gen. Genet. 182, 462–470 (1981). https://doi.org/10.1007/BF00293936
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DOI: https://doi.org/10.1007/BF00293936