Amino acid residues conferring herbicide tolerance in tobacco acetolactate synthase.
Acetolactate synthase (ALS) is the common enzyme in the biosynthetic pathways leading to valine, leucine, and isoleucine in plants and microorganisms. ALS is the target site of several classes of structurally unrelated herbicides including sulfonylureas, imidazolinones, and triazolopyrimidines. To identify the residues conferring herbicide tolerance in tobacco ALS, site-directed mutagenesis for three residues, Ala121, Pro187 and Ser652, was performed. Mutant A121T showed strong resistance to Londax (a sulfonylurea) and Cadre (an imidazolinone), while mutant S652T was resistant only to Cadre. The S652N mutation abolished the binding affinity of FAD, and inactivated the enzyme. Double mutation of Ala121 and Ser652 with Thr yielded a mutant highly tolerant to Londax, Cadre, and TP (a triazolopyrimidine sulfonamide), but has enzymatic properties similar to those of wild-type. Substitution of Pro187 with Ser resulted in the enzyme highly susceptible to oxidation and fragmentation. These results suggest that two residues Ala121 and Ser652 are potent residues conferring herbicide resistance in tobacco ALS, and that double mutation of Ala121 and Ser652 by Thr can confer stronger tolerance to Londax, Cadre, and TP.
Chong
C K
CK
School of Life Science, Cheongju, 361-763, Korea.
Choi
J D
JD
D000095:Acetolactate Synthase / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D000409:Alanine; D019943:Amino Acid Substitution; D004361:Drug Tolerance; D004790:Enzyme Induction; D006540:Herbicides / Q000494:pharmacology*; D007093:Imidazoles / Q000494:pharmacology; D007700:Kinetics; D016297:Mutagenesis, Site-Directed; D010947:Plants, Toxic; D011392:Proline; D011743:Pyrimidines / Q000494:pharmacology; D011994:Recombinant Proteins / Q000737:chemistry / Q000378:metabolism; D012694:Serine; D013329:Structure-Activity Relationship; D013453:Sulfonylurea Compounds / Q000494:pharmacology; D014026:Nicotiana / Q000187:drug effects* / Q000201:enzymology* / Q000235:genetics; D014230:Triazoles / Q000494:pharmacology
D006540:Herbicides; D007093:Imidazoles; D011743:Pyrimidines; D011994:Recombinant Proteins; D013453:Sulfonylurea Compounds; D014230:Triazoles; D012694:Serine; D011392:Proline; D000095:Acetolactate Synthase; D000409:Alanine