Journal Article

Cloning and Characterization of a Hydroxycinnamoyl-CoA:Tyramine <i> N</i>-(Hydroxycinnamoyl)Transferase Induced in Response to UV-C and Wounding from <i>Capsicum annuum</i>

Kyoungwhan Back, Sun Mi Jang, Bong-Choon Lee, Axel Schmidt, Dieter Strack and Kyung-Moon Kim

in Plant and Cell Physiology

Published on behalf of Japanese Society of Plant Physiologists

Volume 42, issue 5, pages 475-481
Published in print May 2001 | ISSN: 0032-0781
Published online May 2001 | e-ISSN: 1471-9053 | DOI: http://dx.doi.org/10.1093/pcp/pce060
Cloning and Characterization of a Hydroxycinnamoyl-CoA:Tyramine
 N-(Hydroxycinnamoyl)Transferase Induced in Response to UV-C and
Wounding from Capsicum
annuum

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  • Biochemistry
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Hydroxycinnamoyl-CoA : tyramine N-(hydroxycinnamoyl) transferase (THT) is a pivotal enzyme in the synthesis of N-(hydroxycinnamoyl)-amines, which are associated with cell wall fortification in plants. The cDNA encoding THT was cloned from the leaves of UV-C treated Capsicum annuum (hot pepper) using a differential screening strategy. The predicted protein encoded by the THT cDNA is 250 amino acids in length and has a relative molecular mass of 28,221. The protein sequence derived from the cDNA shares 76% and 67% identity with the potato and tobacco THT protein sequences, respectively. The recombinant pepper THT enzyme was purified using a bacterial overexpression system. The purified enzyme has a broad substrate specificity including acyl donors such as cinnamoyl-, sinapoyl-, feruloyl-, caffeoyl-, and 4-coumaroyl-CoA and acceptors such as tyramine and octopamine. In UV-C treated plants, the THT mRNA was strongly induced in leaves, and the elevated level of expression was stable for up to 36 h. THT mRNA also increased in leaves that were detached from the plant but not treated with UV-C. THT expression was measured in different plant tissues, and was constitutive at a similar level in leaf, root, stem, flower and fruit. Induction of THT mRNA was correlated with an increase in THT protein.

Keywords: Key words: Capsicum annuum — Cell wall — Hydroxycinnamoyl-CoA:tyramine N-(hydroxycinnamoyl)transferase — UV-C treatment — N-(hydroxycinnamoyl)-amines.; Abbreviations: THT, hydroxycinnamoyl-CoA:tyramine N-(hydroxycin namoyl) transferase; BSA, bovine serum albumin; IPTG, isopropyl β-d-thiogalactopyranoside.; The nucleotide sequence reported in this paper has been submitted to the GenBank under accession number AF329463.

Journal Article.  3970 words.  Illustrated.

Subjects: Biochemistry ; Molecular and Cell Biology ; Plant Sciences and Forestry

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