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K. S. Jakes, in Molecular Action of Toxins and Viruses, P. Cohen and S. van Heyningen, Eds. (Elsevier, Amsterdam, 1982). pp. 131-167; A. P. Pugsley and B. Oudega, in Plasmids: A Practical Approach, K. C. Hardy, Ed. (IRL, Oxford, UK, 1987), pp. 105-161.
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H. Masaki, S. Yajima, A. Akutsu-Koide, T. Ohta, T. Uozumi, in Bacteriocins, Microcins and Lantibiotics, R. James, C. Lazdunski, F. Pattus, Eds. (Springer-Verlag, Berlin, 1992), pp. 379-395; P. C. K. Lau, M. Parsons, T. Uchimura, in Bacteriocins, Microcins and Lantibiotics, R. James, C. Lazdunski, F. Pattus, Eds. (Springer-Verlag, Berlin, 1992), pp. 353-378; R. James, C. Kleanthous, G. R. Moore, Microbiology 142, 1569 (1996).
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Ohta, T.4
Uozumi, T.5
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R. James, C. Lazdunski, F. Pattus, Eds. Springer-Verlag, Berlin
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H. Masaki, S. Yajima, A. Akutsu-Koide, T. Ohta, T. Uozumi, in Bacteriocins, Microcins and Lantibiotics, R. James, C. Lazdunski, F. Pattus, Eds. (Springer-Verlag, Berlin, 1992), pp. 379-395; P. C. K. Lau, M. Parsons, T. Uchimura, in Bacteriocins, Microcins and Lantibiotics, R. James, C. Lazdunski, F. Pattus, Eds. (Springer-Verlag, Berlin, 1992), pp. 353-378; R. James, C. Kleanthous, G. R. Moore, Microbiology 142, 1569 (1996).
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H. Masaki, S. Yajima, A. Akutsu-Koide, T. Ohta, T. Uozumi, in Bacteriocins, Microcins and Lantibiotics, R. James, C. Lazdunski, F. Pattus, Eds. (Springer-Verlag, Berlin, 1992), pp. 379-395; P. C. K. Lau, M. Parsons, T. Uchimura, in Bacteriocins, Microcins and Lantibiotics, R. James, C. Lazdunski, F. Pattus, Eds. (Springer-Verlag, Berlin, 1992), pp. 353-378; R. James, C. Kleanthous, G. R. Moore, Microbiology 142, 1569 (1996).
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James, R.1
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Moore, G.R.3
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The plasmid producing E5-CRD and ImmE5 was constructed as follows. A Nco 1 site (CCATGG) was introduced into the initiation codon of the colE3 gene of pSH357 [H. Masaki, A. Akutsu, T. Uozumi, T. Ohta, Gene 107, 133 (1991)]. and the colE3-immE3 segment from this Nco 1 site to the Bcl 1 (TGATCA) site was replaced with a polymerase chain reaction (PCR)-amplified ColE5-099 DNA fragment, giving rise to plasmid pTO501, which produces the E5-CRD/ImmE5 complex in response to SOS inducers. For this purpose, an Nco 1 site was created at the 5′ end of the PCR fragment in such a way that the Asp at the 116th residue from the COOH-terminus of E5 was changed to an fMet, where the ATG is within the Nco 1 site. The termination codon of immE5 was changed to TGA to create a Bcl 1 site at the 3′ end of the PCR fragment.
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(1991)
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Masaki, H.1
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Uozumi, T.3
Ohta, T.4
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14
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84920313362
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note
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2-terminal sequences of both ImmE5 proteins were identical and lacked fMet. Through similar construction, E3-CRD was produced from the truncated co/E3 gene encoding the COOH-terminal 110 amino acids plus an fMet, and the final product was confirmed to lack fMet.
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16
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84920313361
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data not shown
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T. Ogawa et al., data not shown.
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Ogawa, T.1
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19
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84920313360
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note
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Figure 2B suggests almost the same size for the E3 fragment and the minor RNA band by E5-CRD, but in fact, just the 5′ end was not determined in the sequencing of the E5-CRD minor band (11). The new high-resolution gel suggests that the E5-CRD minor band is 1 base smaller (48 bases?) than the major E3 fragment (49 bases). The 3′ next sequence of the E3-sensitive site, A50-G49, is G49-U48 (numbers being from the 3′ end) in 16S rRNA. If E5-CRD preferentially cleaves this GU in 165 rRNA, this interestingly resembles the cleavage site, GU (QU), within the E5-sensitive tRNA anticodons (Fig. 3B).
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20
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0039525457
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Preliminary data were presented at the 24th Symposium on Nucleic Acids Chemistry (Tokyo): H. Masaki et al., Nucleic Acids Symp. Ser. 37, 287 (1997).
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(1997)
Nucleic Acids Symp. Ser.
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H. Kasai et al., Biochemistry 14, 4198 (1975); S. Noguchi, Y. Nishimura, Y. Hirota, S. Nishimura, J. Biol. Chem. 257, 6544 (1982); S. Nishimura, Prog. Nucleic Add Res. Mol. Biol. 28, 49 (1983).
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H. Kasai et al., Biochemistry 14, 4198 (1975); S. Noguchi, Y. Nishimura, Y. Hirota, S. Nishimura, J. Biol. Chem. 257, 6544 (1982); S. Nishimura, Prog. Nucleic Add Res. Mol. Biol. 28, 49 (1983).
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23
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H. Kasai et al., Biochemistry 14, 4198 (1975); S. Noguchi, Y. Nishimura, Y. Hirota, S. Nishimura, J. Biol. Chem. 257, 6544 (1982); S. Nishimura, Prog. Nucleic Add Res. Mol. Biol. 28, 49 (1983).
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Nishimura, S.1
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24
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84920313359
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-
note
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2. The DNA probe for the 3′ terminal fragment of 16S rRNA was a 23-mer probe.
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27
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84920313358
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note
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Escherichia coli K12 strain JE7338 (F+ tsx purE lac str) and its tgt (TGT-defective) mutant, JE7339, were obtained from the Genetic Stock Research Center, NIG, Japan. A spontaneous btuB (receptor-defective) mutant of JE7338 was isolated with colicin E3, and its resistance to E3, E5, and phage BF23 was confirmed.
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28
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0022429251
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Colicin E3 was purified according to H. Masaki and T. Ohta [J. Mol. Biol. 182, 217 (1985)]. Colicin E5 was partially purified from F. coli K12 RR1(ColE5-099) cells as in the case of the E5-CRD/ImmE5 complex up to the DEAE-TOYOPEARL column chromatography (9). Its purity was densitometrically estimated.
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J. Mol. Biol.
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P. Blackburn and S. Moore, Enzymes 15, 317 (1982); J. Steyaert, K. Hallenga, L. Wyns, P. Stanssens, Biochemistry 29, 9064 (1990).
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(1990)
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J. A. Gerlt, in Nucleases, S. M. Linn, R. S. Lloyd, R. J. Roberts, Eds. (Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY, ed. 2, 1993), pp. 1-34.
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0032522299
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J. Sagara et al., Nucleic Acids Res. 26, 1974 (1998). We recently confirmed that synthetic RNAs devoid of base modifications are sensitive to E5-CRD as well (T. Ogawa et al., in preparation).
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(1998)
Nucleic Acids Res.
, vol.26
, pp. 1974
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Sagara, J.1
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in preparation
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J. Sagara et al., Nucleic Acids Res. 26, 1974 (1998). We recently confirmed that synthetic RNAs devoid of base modifications are sensitive to E5-CRD as well (T. Ogawa et al., in preparation).
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0030592511
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Y. Nakamura, K. Ito, L. A. Isaksson, Cell 87, 147 (1996); K. Ito, M. Uno, Y. Nakamura, Proc. Natl. Acad. Sci. U.S.A. 95, 8165 (1998).
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Y. Nakamura, K. Ito, L. A. Isaksson, Cell 87, 147 (1996); K. Ito, M. Uno, Y. Nakamura, Proc. Natl. Acad. Sci. U.S.A. 95, 8165 (1998).
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Ito, K.1
Uno, M.2
Nakamura, Y.3
-
39
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84920313356
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note
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It was known that there is a great discrepancy between the RNA degradation and ribosorne inactivation activities of the E3-T2A fragment (almost the same as E3-CRD). Usually about 1 μg and 1 ng of E3-T2A were used for RNAs and ribosomes, respectively (12). Because the RNA degradation activity of ES-CRD was in fact higher than that of E3-CRD, the discrepancy of the concentrations used was reduced to about two orders of magnitude.
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40
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0023505590
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Y. Kuchino, N. Hanyu, S. Nishimura, Methods Enzymol. 155, 379 (1987); G. Keith, Biochimie 77, 142 (1995). Samples were developed on Avicel SF glass plates (10 cm by 10 cm) with the following solvent systems: first dimension, isobutyric acid/ammonia/water (66:1:33, v/v/v); second dimension in a and b, isopropanol/concentrated HCl/water (70:15:15, v/v/v); and in c, 100 ml of 0.1 M sodium phosphate buffer (pH 6.8) + 60 g of ammonium sulfate + 2 ml of n-propanol.
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Methods Enzymol.
, vol.155
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Kuchino, Y.1
Hanyu, N.2
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41
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0028936082
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Y. Kuchino, N. Hanyu, S. Nishimura, Methods Enzymol. 155, 379 (1987); G. Keith, Biochimie 77, 142 (1995). Samples were developed on Avicel SF glass plates (10 cm by 10 cm) with the following solvent systems: first dimension, isobutyric acid/ammonia/water (66:1:33, v/v/v); second dimension in a and b, isopropanol/concentrated HCl/water (70:15:15, v/v/v); and in c, 100 ml of 0.1 M sodium phosphate buffer (pH 6.8) + 60 g of ammonium sulfate + 2 ml of n-propanol.
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(1995)
Biochimie
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-
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Keith, G.1
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43
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84920313355
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note
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We thank I. Nitta and M. Sano for technical advice; M. Hidaka, A. Nakamura, Y. Gunji, and J. Sagara for helpful discussions; and D. P. Brunner for ColE5-099. This work was supported by a Grant-in-Aid for Scientific Research from the Ministry of Education, Science, Sports and Culture.
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