A nuclear mutation conferring thiostrepton resistance in Chlamydomonas reinhardtii affects a chloroplast ribosomal protein related to Escherichia coli ribosomal protein L11.
We have isolated a nuclear mutant (tsp-1) of Chlamydomonas reinhardtii which is resistant to thiostrepton, an antibiotic that blocks bacterial protein synthesis. The tsp-1 mutant grows slowly in the presence or absence of thiostrepton, and its chloroplast ribosomes, although resistant to the drug, are less active than chloroplast ribosomes from the wild type. Chloroplast ribosomal protein L-23 was not detected on stained gels or immunoblots of total large subunit proteins from tsp-1 probed with antibody to the wild-type L-23 protein from C. reinhardtii. Immunoprecipitation of proteins from pulse-labeled cells showed that tsp-1 synthesizes small amounts of L-23 and that the mutant protein is stable during a 90 min chase. Therefore the tsp-1 phenotype is best explained by assuming that the mutant protein synthesized is unable to assemble into the large subunit of the chloroplast ribosome and hence is degraded over time. L-23 antibodies cross-react with Escherichia coli r-protein L11, which is known to be a component of the GTPase center of the 50S ribosomal subunit. Thiostrepton-resistant mutants of Bacillus megaterium and B. subtilis lack L11, show reduced ribosome activity, and have slow growth rates. Similarities between the thiostrepton-resistant mutants of bacteria and C. reinhardtii and the immunological relatedness of Chlamydomonas L-23 to E. coli L11 suggest that L-23 is functionally homologous to the bacterial r-protein L11.
McElwain
K B
KB
Department of Zoology, Duke University, Durham, NC 27708-0325.
Boynton
J E
JE
Gillham
N W
NW
D000818:Animals; D015153:Blotting, Western; D016825:Chlamydomonas reinhardtii / Q000187:drug effects*; D002736:Chloroplasts / Q000737:chemistry*; D015180:Electrophoresis, Gel, Two-Dimensional; D029968:Escherichia coli Proteins; D009154:Mutation; D011233:Precipitin Tests; D012269:Ribosomal Proteins / Q000235:genetics* / Q000276:immunology; D012270:Ribosomes / Q000737:chemistry / Q000187:drug effects*; D013883:Thiostrepton / Q000494:pharmacology*
D029968:Escherichia coli Proteins; D012269:Ribosomal Proteins; C027624:ribosomal protein L11; C043816:ribosomal protein L23, E coli; D013883:Thiostrepton