Publications

• Biosynthesis and accumulation of sterols.

  Benveniste P.

  In recent years, the impressive development of molecular genetics tools, the sequencing of the Arabidopsis thaliana genome, the availability of DNA or transposon tagged mutants, and the multiple possibilities offered by stable transformation with DNA in sense and antisense orientation have enabled ... >> More

  In recent years, the impressive development of molecular genetics tools, the sequencing of the Arabidopsis thaliana genome, the availability of DNA or transposon tagged mutants, and the multiple possibilities offered by stable transformation with DNA in sense and antisense orientation have enabled the application of a strategy of gain or loss of function to study the sterol biosynthesis pathway. Here we describe the results obtained with these techniques. The results essentially confirm data obtained previously with sterol biosynthesis inhibitors (SBIs) and enable the precise dissection of biosynthetic pathways. We discuss the advantages and disadvantages of molecular genetics techniques as applied to sterol metabolism. The greater selectivity of these techniques constitutes an invaluable advantage and has led to the discovery of a role for sterols in plant development. << Less

  Annu Rev Plant Biol 55:429-457(2004) [PubMed] [EuropePMC]

• Sterol composition and growth of transgenic tobacco plants expressing type-1 and type-2 sterol methyltransferases.

  Sitbon F., Jonsson L.

  Transgenic tobacco (Nicotiana tabacum L.) plants with altered sterol composition were generated by transformation with plant cDNAs encoding type-1 and type-2 sterol methyltransferases (SMTs; EC 2.1.1.41). For both SMT1 and SMT2 transformants, the transformation was associated with a reduction in t ... >> More

  Transgenic tobacco (Nicotiana tabacum L.) plants with altered sterol composition were generated by transformation with plant cDNAs encoding type-1 and type-2 sterol methyltransferases (SMTs; EC 2.1.1.41). For both SMT1 and SMT2 transformants, the transformation was associated with a reduction in the level of cholesterol, a non-alkylated sterol. In SMT1 transformants a corresponding increase of alkylated sterols, mainly 24-methyl cholesterol, was observed. On the other hand, in SMT2 transformants the level of 24-methyl cholesterol was reduced, whereas the level of sitosterol was raised. No appreciable alteration of total sterol content was observed for either genotype. The general phenotype of transformants was similar to that of controls, although SMT2 transformants displayed a reduced height at anthesis. The results show that plant sterol composition can be altered by transformation with an SMT1 cDNA without adverse effects on growth and development, and provide evidence, in planta, that SMT1 acts at the initial step in sterol alkylation. << Less

  Planta 212:568-572(2001) [PubMed] [EuropePMC]

• Two families of sterol methyltransferases are involved in the first and the second methylation steps of plant sterol biosynthesis.

  Bouvier-Nave P., Husselstein T., Benveniste P.

  Two methyl transfers are involved in the biosynthesis of 24-methyl and 24-ethyl sterols, which play major roles in plant growth and development. The first methyl transfer applies to cycloartenol, the second to 24-methylene lophenol. About ten cDNA clones encoding S-adenosyl-L-methionine (AdoMet) s ... >> More

  Two methyl transfers are involved in the biosynthesis of 24-methyl and 24-ethyl sterols, which play major roles in plant growth and development. The first methyl transfer applies to cycloartenol, the second to 24-methylene lophenol. About ten cDNA clones encoding S-adenosyl-L-methionine (AdoMet) sterol methyltransferases (SMTs) have been isolated so far from various plants. According to their deduced amino acid sequences, they were classified in two families, smtl and smt2; in addition, smt2 cDNAs were shown to encode a 24-methylene lophenol C24 methyltransferase [Bouvier-Navé, P., Husselstein, T., Desprez, T. & Benveniste, P. (1997) Eur. J. Biochem. 246, 518-529]. We now report the comparison of two cDNAs isolated from Nicotiana tabacum, Ntsmt1-1 which belongs to the first SMT cDNA family and Ntsmt2-1 which belongs to the second. Both cDNAs were expressed in the yeast null mutant erg6, deficient in SMT. Whereas erg6 is devoid of 24-alkyl sterols, erg6 Ntsmt1-1 contained a majority of 24-methylene sterols and erg6 Ntsmt2-1, a majority of 24-ethylidene sterols, indicating distinct functions for the expression products of these cDNAs. In the presence of AdoMet, delipidated microsomes from erg6 Ntsm1-1 efficiently converted cycloartenol into 24-methylene cycloartanol, but did not produce any 24-ethylidene lophenol upon incubation with 24-methylene lophenol. This demonstrates that cDNA Ntsmt1-1 (and most probably the other plant SMT cDNAs of the first family) encode(s) a cycloartenol C24 methyltransferase. In contrast, delipidated microsomes of erg6 Ntsmt2-1 were shown to methylate preferentially 24-methylene lophenol, as expected from an SMT encoded by an smt2 cDNA. In summary, among various cDNAs isolated from N. tabacum, one (Ntsmt1-1) belongs to the first family of plant SMT cDNAs according to its deduced amino acid sequence and was shown to encode a cycloartenol C24 methyltransferase, whereas another (Ntsmt2-1) belongs to the second family and was shown to encode a 24-methylene lophenol C24 methyltransferase. Meanwhile, two cDNAs were isolated from Oriza sativa and shown to belong to smtl and to smt2 families, respectively. These data disclose the coexistence, in a given plant species, of two distinct SMTs, each catalyzing one step of methylation in the sterol biosynthesis pathway. << Less

  Eur. J. Biochem. 256:88-96(1998) [PubMed] [EuropePMC]

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