.=) 1992 Oxford University Press
DNA
Nucleic Acids Research, Vol. 20, No. 6 1423
of the 3' end of the Caulobacter crescentus 16S rRNA gene sequence
B.Ely Department of Biological Sciences, University of South Carolina, Columbia, SC 29208, USA Submitted December 30, 1991 During the course of our analyses of the DNA sequences of Caulobacter crescentus genes, we became suspicious that there might be errors in the published DNA sequence of the 16S ribosomal RNA gene (1). Since newer technology allows more accurate DNA sequence determination, we redetermined the DNA sequence of the 3'-end of the 16S ribosomal RNA genes of C. crescentus using modifications of the dideoxy sequencing procedure which have proved successful for sequence determination of C. crescentus DNA in our laboratory (2,3). After determining the DNA sequence from the XhoI site near the 3'-end of the 16S rRNA genes to the BglI site between the tRNAile gene and the tRNAala gene, we found that both of the 16S ribosomal RNA genes of C. crescentus strain CB15 (4) have identical sequences at the 3'-end. Furthermore, these sequences are identical to that we obtained from the 16S ribsomal RNA gene contained in the C. crescentus CB13 clone used in the earlier sequence determination (1). The differnces between the DNA sequence we report and that of the earlier publication are significant, especially since the sequences of the region involved in the binding of the ribosome to mRNA are different. We have found the identical sequence in three different cloned genes, and this sequence contains a region identical to the RNA sequence originally obtained by Shine and Dalgamo (5) when they determined the sequence of the 16S ribosonial RNA isolated from C. crescentus ribosomes. Thus, we predict that ribosome binding sites found in C. crescentus mRNAs will be complementary to the sequence CCTCCTTTCT. Further, we have demonstrated that identical copies of the tRNAile genes are located downstream from each copy of the 16S rRNA genes.
ACKNOWLEDGEMENTS I thank Lilly Gallman and Nicole Matthews for expert technical assistance. This work was supported by GM34765 from National Institutes of Health.
EMBL accession no. X63362
REFERENCES 1. Feingold,J., Bellafatto,V., Shapiro,L. and Amemiya,K. (1985) J. Bacteriol. 163, 155-166. 2. Schoenlein,P.V., Gallman,L.M. and Ely,B. (1988) Gene 70, 321-329. 3. Khambaty,F. and Ely,B. (1990) BioTechniques 9, 714-715. 4. Ohta,N. and Newton,A. (1981) J. Mol. Biol. 153, 281-303. 5. Shine,J. and Dalgamo,L. (1975) Nature 254, 34-38.
G
A
T
C
T T
T C
C T
C C A
C T A
G
G
Figure 1. The nucleotide sequence of the 3' end of the rrnA 16S rRNA gene from the C. crescentus wild type strain CB15. The EMBL accession number for this sequence is X63362. The corresponding sequence from the rmnB is identical
(X63363).
DNA
Nucleic Acids Research, Vol. 20, No. 6 1423
of the 3' end of the Caulobacter crescentus 16S rRNA gene sequence
B.Ely Department of Biological Sciences, University of South Carolina, Columbia, SC 29208, USA Submitted December 30, 1991 During the course of our analyses of the DNA sequences of Caulobacter crescentus genes, we became suspicious that there might be errors in the published DNA sequence of the 16S ribosomal RNA gene (1). Since newer technology allows more accurate DNA sequence determination, we redetermined the DNA sequence of the 3'-end of the 16S ribosomal RNA genes of C. crescentus using modifications of the dideoxy sequencing procedure which have proved successful for sequence determination of C. crescentus DNA in our laboratory (2,3). After determining the DNA sequence from the XhoI site near the 3'-end of the 16S rRNA genes to the BglI site between the tRNAile gene and the tRNAala gene, we found that both of the 16S ribosomal RNA genes of C. crescentus strain CB15 (4) have identical sequences at the 3'-end. Furthermore, these sequences are identical to that we obtained from the 16S ribsomal RNA gene contained in the C. crescentus CB13 clone used in the earlier sequence determination (1). The differnces between the DNA sequence we report and that of the earlier publication are significant, especially since the sequences of the region involved in the binding of the ribosome to mRNA are different. We have found the identical sequence in three different cloned genes, and this sequence contains a region identical to the RNA sequence originally obtained by Shine and Dalgamo (5) when they determined the sequence of the 16S ribosonial RNA isolated from C. crescentus ribosomes. Thus, we predict that ribosome binding sites found in C. crescentus mRNAs will be complementary to the sequence CCTCCTTTCT. Further, we have demonstrated that identical copies of the tRNAile genes are located downstream from each copy of the 16S rRNA genes.
ACKNOWLEDGEMENTS I thank Lilly Gallman and Nicole Matthews for expert technical assistance. This work was supported by GM34765 from National Institutes of Health.
EMBL accession no. X63362
REFERENCES 1. Feingold,J., Bellafatto,V., Shapiro,L. and Amemiya,K. (1985) J. Bacteriol. 163, 155-166. 2. Schoenlein,P.V., Gallman,L.M. and Ely,B. (1988) Gene 70, 321-329. 3. Khambaty,F. and Ely,B. (1990) BioTechniques 9, 714-715. 4. Ohta,N. and Newton,A. (1981) J. Mol. Biol. 153, 281-303. 5. Shine,J. and Dalgamo,L. (1975) Nature 254, 34-38.
G
A
T
C
T T
T C
C T
C C A
C T A
G
G
Figure 1. The nucleotide sequence of the 3' end of the rrnA 16S rRNA gene from the C. crescentus wild type strain CB15. The EMBL accession number for this sequence is X63362. The corresponding sequence from the rmnB is identical
(X63363).
