Expression and functional assembly into bacterial ribosomes of a nuclear-encoded chloroplast ribosomal protein with a long NH2-terminal extension.
Chloroplast ribosomal protein L13 is encoded in the plant nucleus and is considerably larger than its eubacterial homologue by having NH2- and COOH-terminal extensions with no homology to any known sequences (Phua et al., J Biol. Chem. 264, 1968-1971, 1989). We made two gene constructs of L13 cDNA using the polymerase chain reaction (PCR) and expressed them in Escherichia coli. Analysis of the ribosomes and polysomes from these cells, using an antiserum specific to chloroplast L13, shows that the expressed proteins are incorporated, in the presence of the homologous E. coli L13, into functional ribosomes which participate in protein synthesis (i.e. polysomes). Evidence is obtained that the large NH2-terminal extension probably lies on the surface of these 'mosaic ribosomes.' This first report of the assembly into E. coli ribosomes of nuclear-coded chloroplast ribosomal protein with terminal extensions thus suggest an extraordinary conservation in the function of eubacterial type ribosomal proteins, despite the many changes in protein structure during their evolution inside a eukaryotic system.
Giese
K
K
Max-Planck-Institut für Molekulare Genetik, Abteilung Wittmann, Berlin, Germany.
Subramanian
A R
AR
D000595:Amino Acid Sequence; D001483:Base Sequence; D002736:Chloroplasts / Q000378:metabolism*; D004926:Escherichia coli / Q000378:metabolism*; D015870:Gene Expression / Q000502:physiology; D046911:Macromolecular Substances; D008969:Molecular Sequence Data; D010944:Plants / Q000235:genetics*; D016133:Polymerase Chain Reaction; D011132:Polyribosomes / Q000378:metabolism; D011994:Recombinant Proteins / Q000096:biosynthesis; D012269:Ribosomal Proteins / Q000235:genetics / Q000378:metabolism*; D012270:Ribosomes / Q000378:metabolism* / Q000502:physiology
D046911:Macromolecular Substances; D011994:Recombinant Proteins; D012269:Ribosomal Proteins