Heavy-metal-responsive genes in maize: identification and comparison of their expression upon various forms of abiotic stress.

To identify genes involved in defense against heavy-metal stresses, a cDNA library originating from mercuric chloride-treated maize (Zea mays L. cv. INRA 258) leaves was constructed and analysed by differential screening using cDNAs derived from treated and untreated plants. Transcriptionally activated cDNA clones, designated CHEM (chemically-activated), were isolated and characterized. They represent various known proteins, such as glycine-rich proteins, pathogenesis-related proteins, chaperones and membrane proteins. The expression of the genes encoding these proteins was studied in maize subjected to other forms of abiotic stress. Expression of glycine-rich proteins was greatly enhanced by heat stress, and also stimulated by NaCl, polluted rainwater, wounding and cold stress. Pathogenesis-related proteins were strongly induced by ultraviolet light and to a lesser extent by NaCl, polluted rainwater and wounding. Heat-shock protein was mainly induced by heat and cold, and ubiquitin by wounding. Expression of the membrane channel protein was stimulated by heat stress, NaCl, polluted rainwater and ultraviolet-light irradiation.

The complete amino acid sequence of the pathogenesis-related (PR2) protein induced in chemically stressed bean leaves.

We have established the complete sequence of the 155 amino acid residues of the pathogenesis-related protein PR2 accumulating in bean leaves treated with a mercuric chloride solution. Bean PR2 whose biological function remains to be elucidated, represents a structurally unfamiliar protein in which sequence arginine, cysteine, methionine and tryptophan residues are missing. This sequence is identical to that of bean PvPR1.

The steroid antagonist RU38486 is metabolized by the liver microsomal P450 mono-oxygenases.

Microsomal preparations from adult male rat liver actively oxidized RU38486 into the 11 beta-monodemethylated, 11 beta-didemethylated and 17 alpha-hydroxylated derivatives, metabolites which are known to be formed in vivo. These oxidative reactions were inhibited at different degrees by P450 chemical inhibitors. Pretreatment of the animals by P450 mono-oxygenase prototype inducers led to drastic changes in RU38486 metabolization. Methylcholanthrene treatment carried out a significant decrease while phenobarbital markedly increased the metabolic activity of the liver microsomes. Moreover, antibodies to methylcholantrene-inducible P450 forms did not affect the metabolic activity while a complete blockade-of RU38486 oxidation was observed in the presence of antibodies to phenobarbital- inducible forms. The present results demonstrate that liver P450 mono-oxygenases are engaged in different oxidative steps of RU38486 metabolism and that phenobarbital-inducible but not methylcholanthrene-inducible P450 forms are active in RU38486 degradation.

Structural and immunochemical properties of rat liver tyrosine aminotransferase.

Our results show for the first time sequence data of the N-terminal part of tyrosine aminotransferase. This unblocked form of TATase, which has never been detected before, starts with a serine. This serine was found at position 29 of the primary structure of the enzyme deduced from the cDNA. We suggest that this free N-terminal amino acid is the extremity of a TATase form generated by proteolysis during the process of purification. Thus, proteolysis does not occur at the C-terminal as has been suggested before, but rather at the N-terminal region of the enzyme. To confirm this possibility, a peptide corresponding to the sequence of the seven carboxy-terminal amino acids of TATase was synthesized. It was coupled to ovalbumin or keyhole limpet hemocyanin, and the resulting conjugates were used to raise anti-peptide antibodies. The crude sera obtained were purified and their abilities to recognize TATase in ELISA and dot-blot experiments were proven. Our results demonstrate that the C-terminal part of the enzyme is present and well-recognized by the anti-peptide serum prepared. Furthermore, the anti-peptide serum reacts with TATase without inhibiting its enzymatic activity.

Integrity of the C-terminal part of tyrosine aminotransferase revealed by an anti-peptide serum.

A synthetic peptide corresponding to the seven carboxy-terminal amino acids of tyrosine aminotransferase was coupled to ovalbumin or keyhole limpet haemocyanin, and the resulting conjugates were used to raise anti-peptide antibodies by immunization of rabbits. The crude sera were purified and tested for recognition of the whole enzyme by enzyme-linked immunosorbent assay and immunoprecipitation in extracts from [35S] methionine labeled hepatoma cells. Our results support the existence of an intact C-terminus. If processing takes place, it will rather occur at the N-terminus of this hepatic enzyme.

Effects of 5-HT serotonin on spontaneous locomotor activity of eels.

The swimming activity of eels maintained in tap water at 8-12 degrees C is significantly decreased after i.p. para-chlorophenylalanine (pCPA) administration at dose of 200 mg/kg (p less than 0.005 to respective control value). 5-Hydroxytryptophan (5-HTP) restored transitory swimming activity of eels. These results suggest that 5-HT has an important part in locomotor activity of eels.