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Nucleic Acids Research, 1986, Vol. 14, No. 2 883-898
© 1986

Articles

The enigma of the gene coding for ribosomal protein S12 in the chtoroplasts of Nicotiana

Hillel Fromm, Marvin Edelman, Barbara Koller1, Pierre Goloubinoff and Esra Galun

Department of Plant Genetics, Weizmann Institute of Science Rehovot, Israel 1European Molecular Biology Laboratory Heidelberg, FRG

Received July 2, 1985. Accepted December 10, 1985.

A 2.9 kbp region from within the inverted repeat of Nicotiana chloroplast DNA hybridized with a chloroplast DNA fragment from Euglena containing the complete rps12 gene coding for ribosomal protein S12. Nuoleotide sequencing within this region revealed the existence of two rps12 coding stretches interrupted by 510 bp having class II intron structure. Joining and decoding the exon regions produced a sequence of 85 amino acids colinear and 81% homologous to the S12 protein of Euglena chloroplasts and E. coli, starting from amino acid residue 38 to the stop codon. Immediately upstream of codon 38, conserved intron sequences were located. However, the 5' 37 codons of Nicotiana chloroplast rps12 could not be identified by electron microscopy of RNA-DNA hybrids within a DNA region extending 1000 bp upstream of codon 38, nor by computer search of a completely sequenced region extending for more than 9000 bp upstream of this codon. In E. coli, alteration in rps12 codons 42 or 87 causes streptomycin resistance. However, the nucleotide sequence of the identified rps12 exons in two Nicotiana chloroplast mutants resistant to streptomycin were found to be identical to that of wild type.


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