Phenylalanine ammonia-lyase in tobacco mosaic virus-infected hypersensitive tobacco. Density-labelling evidence of de novo synthesis.
A strong increase in phenylalanine ammonia-lyase (EC 4.3.1.5) activity occurs in tobacco mosaic virus-infected tobacco leaves developing necrotic local lesions. Comparison of physicochemical properties of the partially purified enzymes extracted from healthy and infected leaves showed that the hypersensitive reaction leads to an increase in the pool size of the same active enzyme molecules as those present in non-infected material. The molecular mechanism of enzyme formation was investigated by radiolabelling with [3H]leucine and by density labelling with 2H2O. Abnormal patterns of incorporation of radioactivity into all soluble proteins were found in infected leaves carrying local lesions. In contrast, uptake of deuterium into the amino acid pool was the same in healthy and infected leaves. Unstimulated phenylalanine ammonia-lyase was shown to be a long-lived enzyme (half-life: 25-35 h). Results of comparative density labelling experiments unequivocally demonstrated that the increased enzyme pool size arose from an increased rate of synthesis mediated by the hypersensitive reaction.
D000135:Acid Phosphatase / Q000096:biosynthesis; D000642:Ammonia-Lyases / Q000096:biosynthesis*; D003903:Deuterium; D007553:Isotope Labeling; D007700:Kinetics; D010650:Phenylalanine Ammonia-Lyase / Q000096:biosynthesis* / Q000302:isolation & purification; D010944:Plants / Q000201:enzymology; D010947:Plants, Toxic; D014026:Nicotiana / Q000201:enzymology; D014027:Tobacco Mosaic Virus / Q000201:enzymology*; D014779:Virus Replication; D001616:beta-Galactosidase / Q000096:biosynthesis
D003903:Deuterium; D000135:Acid Phosphatase; D001616:beta-Galactosidase; D000642:Ammonia-Lyases; D010650:Phenylalanine Ammonia-Lyase