Molecular characterization of genes coding for wild-type and sulfonylurea-resistant acetolactate synthase in the cyanobacterium Synechococcus PCC7942.
We present here the isolation and molecular characterization of acetolactate synthase (ALS) genes from the cyanobacterium Synechococcus PCC7942 which specify a sulfonylurea-sensitive enzyme and from the sulfonylurea-resistant mutant SM 3/20, which specify resistance to sulfonylurea herbicides. The ALS gene was cloned and mapped by complementation of an Escherichia coli ilv auxotroph that requires branched-chain amino acids for growth and lacks ALS activity. The cyanobacterial gene is efficiently expressed in this heterologous host. The ALS gene codes for 612 amino acids and shows high sequence homology (46%) at the amino acid level with ALS III of E. coli and with the tobacco ALS. The resistant phenotype is a consequence of proline to serine substitution in residue 115 of the deduced amino acid sequence. Functional expression of the mutant gene in wild-type Synechococcus and in E. coli confirmed that this amino-acid substitution is responsible for the resistance. Yet the deduced amino-acid sequence as compared with other ALS proteins supports the notion that the amino-acid context of the substitution is important for the resistance.
Friedberg
D
D
Division of Microbial and Molecular Ecology, Hebrew University of Jerusalem, Israel.
Seijffers
J
J
D000095:Acetolactate Synthase / Q000235:genetics*; D000595:Amino Acid Sequence; D001483:Base Sequence; D015139:Blotting, Southern; D003001:Cloning, Molecular; D000458:Cyanobacteria / Q000187:drug effects / Q000201:enzymology / Q000235:genetics*; D004247:DNA / Q000235:genetics / Q000302:isolation & purification; D004351:Drug Resistance / Q000235:genetics; D017343:Genes, Plant; D006540:Herbicides / Q000494:pharmacology*; D008969:Molecular Sequence Data; D009154:Mutation; D007652:Oxo-Acid-Lyases / Q000235:genetics*; D015183:Restriction Mapping; D012689:Sequence Homology, Nucleic Acid; D013453:Sulfonylurea Compounds / Q000494:pharmacology*
D006540:Herbicides; D013453:Sulfonylurea Compounds; D004247:DNA; D000095:Acetolactate Synthase; D007652:Oxo-Acid-Lyases