3514 Nucleic Acids Research, Vol. 20, No. 13
.=) 1992 Oxford University Press
Nucleotide sequence of tryptophan tRNA from Bacillus subtilis Jitsuhiro Matsugi, Katsutoshi Murao and Hisayuki Ishikura 331 1-1, Laboratory of Chemistry, Jichi Medical School, Minamikawachi-machi, Tochigi-ken, 329-04, Japan Submitted May 26, 1992 B.subtilis tryptophan tRNA was isolated utilizing a dot blot hybridization method (1) and its RNA sequence including a modified nucleotide was determined by 2D TLC (2, 3) and UV spectroscopy (4). As a result, we identified five modified nucleotides, (ribothymidine (T54), pseudouridine (*55, *31), M-isopentenyladenosine (i6A37) and dihydrouridine (D20)). In B.subtilis, it has been re d tt a UGA termination codon was translated as tryptophan at low efficiency (5). However, a single tryptophan tRNA gene has been reported so far by Wawrousek et al. (6) and consequently, the anticodon sequence of the tRNA corresponding to the UGA codon comes into question. In this regard, we found that this tRNA has the anticodon 5'CCA3' and no modification in the first letter of the anticodon. The question whether this tryptophan tRNA reads a UGA codon like that of E. coli (7) or B.subtilis has another unknown isoacceptor is now under investigation.
REFERENCES 1. Yokogawa,T., Kumazawa,Y., Miura,K. and Watanabe,K. (1989) Nucleic Acids Res. 17, 2623-2638. 2. Kimura,F., Harada,F. and Nishimura,S. (1971) Biochemistry, 10, 3277-3283. 3. Silberklang,M., Prochiantz,A., Haenni,A.L. and RajBhandary,U.L. (1977) Eur. J. Biochem. 72, 465-478. 4. Hall,R.H. (1971) 7he Modified Nucleotides in Nucleic Acids, Columbia University Press, New York, London. 5. Lovett,P.S., Ambulos,N.P.,Jr, Mulbry,W., Noguchi,N. and Rogers,E.J. (1991) J. Bacteriol. 173, 1810-1812.
DDBJ accession no. D10981 6. Wawrousek,E.F., Narasimhan,N. and Hansen,J.N. (1984) J. Biol. Chem. 259, 3694-3702. 7. Hirsh,D. (1971) J. Mol. Biol. 58, 439-458.
A C C G G *C G *C G *C G *C C *G UAt ~~~~~
A uu A CAIJCCU C AAUJUUGAU G 0o...o *0** G G UG C G DAGAACA GvCC GG 20 5 20 ~~~AoU G *C G *C 311v*A C A U i1A37
T50
CcA
Figure. Secondary stnicture of B.subtilis tryptophan tRNA.
.=) 1992 Oxford University Press
Nucleotide sequence of tryptophan tRNA from Bacillus subtilis Jitsuhiro Matsugi, Katsutoshi Murao and Hisayuki Ishikura 331 1-1, Laboratory of Chemistry, Jichi Medical School, Minamikawachi-machi, Tochigi-ken, 329-04, Japan Submitted May 26, 1992 B.subtilis tryptophan tRNA was isolated utilizing a dot blot hybridization method (1) and its RNA sequence including a modified nucleotide was determined by 2D TLC (2, 3) and UV spectroscopy (4). As a result, we identified five modified nucleotides, (ribothymidine (T54), pseudouridine (*55, *31), M-isopentenyladenosine (i6A37) and dihydrouridine (D20)). In B.subtilis, it has been re d tt a UGA termination codon was translated as tryptophan at low efficiency (5). However, a single tryptophan tRNA gene has been reported so far by Wawrousek et al. (6) and consequently, the anticodon sequence of the tRNA corresponding to the UGA codon comes into question. In this regard, we found that this tRNA has the anticodon 5'CCA3' and no modification in the first letter of the anticodon. The question whether this tryptophan tRNA reads a UGA codon like that of E. coli (7) or B.subtilis has another unknown isoacceptor is now under investigation.
REFERENCES 1. Yokogawa,T., Kumazawa,Y., Miura,K. and Watanabe,K. (1989) Nucleic Acids Res. 17, 2623-2638. 2. Kimura,F., Harada,F. and Nishimura,S. (1971) Biochemistry, 10, 3277-3283. 3. Silberklang,M., Prochiantz,A., Haenni,A.L. and RajBhandary,U.L. (1977) Eur. J. Biochem. 72, 465-478. 4. Hall,R.H. (1971) 7he Modified Nucleotides in Nucleic Acids, Columbia University Press, New York, London. 5. Lovett,P.S., Ambulos,N.P.,Jr, Mulbry,W., Noguchi,N. and Rogers,E.J. (1991) J. Bacteriol. 173, 1810-1812.
DDBJ accession no. D10981 6. Wawrousek,E.F., Narasimhan,N. and Hansen,J.N. (1984) J. Biol. Chem. 259, 3694-3702. 7. Hirsh,D. (1971) J. Mol. Biol. 58, 439-458.
A C C G G *C G *C G *C G *C C *G UAt ~~~~~
A uu A CAIJCCU C AAUJUUGAU G 0o...o *0** G G UG C G DAGAACA GvCC GG 20 5 20 ~~~AoU G *C G *C 311v*A C A U i1A37
T50
CcA
Figure. Secondary stnicture of B.subtilis tryptophan tRNA.
