Chloroplast ribosomal protein L13 is encoded in the nucleus and is considerably larger than its bacterial homologue. Construction, immunoisolation, and nucleotide sequence (including transit peptide) its cDNA clone from an angiosperm.
Chloroplast ribosomes of higher plants are of the prokaryotic ribosome motif but, unlike in bacteria, their ribosomal protein (r-protein) genes are distributed between the organelle and the nucleus. In order to isolate some of the nuclear-encoded r-protein genes, we have raised antibodies to several spinach chloroplast r-proteins and constructed spinach cDNA expression libraries in lambdagt11. Screening the libraries with one of the antisera yielded three cDNA clones for r-protein L13, an early 50 S subunit assembly protein essential for RI50 formation. The cDNA clone encodes, beginning with a Met codon in the consensus plant initiator context, a polypeptide of 250 amino acid residues. The NH2-terminal 60 residues bear the characteristic features of a chloroplast transit peptide. The putative mature L13 protein, which has common immunoepitopes with Escherichia coli L13, is 34% longer than the E. coli homologue. It has 56% sequence identity with E. coli L13 in the homologous region, but no identity to any known protein in the extra stretch. There are two neighboring ATG codons in the 5' region and two putative plant polyadenylation signals in the 3'-untranslated region of the cDNA. Their possible effect to increase translational efficiency is discussed, and the importance of encoding a RI50 protein in the nuclear genome for possible nuclear control of chloroplast protein synthesis is noted.
Phua
S H
SH
Max-Planck-Institut für Molekulare Genetik, Abt. Wittmann, Berlin, Federal Republic of Germany.
Srinivasa
B R
BR
Subramanian
A R
AR
D000595:Amino Acid Sequence; D001483:Base Sequence; D002467:Cell Nucleus / Q000378:metabolism*; D002736:Chloroplasts / Q000378:metabolism*; D004926:Escherichia coli / Q000235:genetics*; D005796:Genes; D008969:Molecular Sequence Data; D010944:Plants / Q000235:genetics*; D015183:Restriction Mapping; D012269:Ribosomal Proteins / Q000235:genetics*; D012689:Sequence Homology, Nucleic Acid
D012269:Ribosomal Proteins