k. 1991 Oxford University Press
2778 Nucleic Acids Research, Vol. 19, No. 10
Nicolas Biteau*, Christophe Fremaux, Sylvie Hebrard, Michel Aigle and Marc Crouzet Laboratoire de Biologie Moleculaire et de Sequen9age, Universite de Bordeaux 11, 146 rue Leo Saignat, 33076 Bordeaux Cedex, France EMBL accession no. X58787
Submitted April 9, 1991 As a part of the European program BAP to sequence the chromosome El[ of Saccharomyces cerevisiae (strain J24-24a), we have determined the complete nucleotide sequence of a 10.85 kb BamHI DNA fragment located to the right of the centromere. This region reveals the presence of two tRNA genes. One codes for the tRNAPro carrying the 5'-AGG-3' anticodon. The gene was already identified by suppression of fiameshift mutations and was named SUF2 (1). The gene was cloned and the nucleic acid sequence of the region carrying SUF2 was published (2). No nucleotide difference was observed with our results. The computer analysis of the BamHI DNA fragment has shown that the other gene encodes a tRNAAsn carrying the 5'-GTT-3' anticodon (Fig. lA). The tRNAAsn was the only amino acid acceptor for which DNA sequence data were lacking (3). Nevertheless this tRNA was purified and the ribonucleic sequence was determined using post-labeling methods (4). The sequence deduced from tRNAAsn compared to the corresponding gene showed that the gene is colinear with the mature product. The gene does not contain any intervening sequences. As expected in eucaryotes (3), the 3'-CCA nucleotides found at the termini of all tRNAs are not encoded by the gene. The cloverleaf structure of this tRNA without the post-transcriptional modifications is represented in the figure 1B.
3. Guthrie,C. and Abelson,J. (1982) The molecular biology of the yeast Saccharomyces (eds, N.Strathem, E.Jones and J.Broach), 487-528. 4. Keith,G. and Pixa,G. (1984) Biochimie 66, 639-643
A 50 40 10 30 20 I I 1 TCAACATATATTAGTAATACTTTTTCTGTAAAAGTGACACAAGATAAAAT
101 TGATTCAACCCACT GGATi2, TTAATTTTTCTTTTCTTTTTTTTTTT 151 TTATCTCCGACGGAGAAAAAATTT
Figure 1A. Nucleotide sequence of the region containing the tRNAAsn gene. The sequence for tRNAASn is underlined.
G 1.G - C A -T C - G. T G 70 5.C - G C- G C C A -T C A C T C T T G T A G G T G A G A C C G
A G G C
T A A A G
TG - C
30 C - G
REFERENCES 1. Cumniins,C.M., Gaber,R.F., Culbertson,M.R., Mann,R. and Fink,G.R. (1980) Genetics 95, 855-875. 2. Cumniins,C.M., Donahue,T.F. and Culbertson M.R. (1982) Proc. Natl. Acad. Sci. USA 79, 3565-3569.
To whom correspondence should be addressed
.G - C. A - T 40 A A T G T
Figure 1B. Cloverleaf structure of tRNAAS" as predicted by informatic analysis.