Molecular characterization of an h-type thioredoxin gene from grape

Abstract

Abstract Thioredoxins (Trxs), as small ubiquitous proteins, participate in dithiol-disulfide exchange reactions. In contrast to other organisms, plants have a complex family of Trxs, which contains seven different Trx types: f, h, m, o, x, y, and z. The h-type Trx consists of multiple forms that are involved in different processes. A full-length cDNA coding for a Trx h, designated VvTrx h2, was isolated and cloned from grape (Vitis vinifera L. cv. White Seedless) berry tissue by RT-PCR technique. Nucleotide sequence analysis revealed 561 nucleotides in length encoded for a protein of 114 amino acid residues. The deduced polypeptide sequence harbors a typical catalytic site, WCGPC and its calculated molecular mass and its predicted isoelectric point are 12.79 and 5.06 kDa, respectively. The threedimensional modeling and docking studies allow for the proposal that VvTrx h2 could be reduced by a NADP-thioredoxin reductase rather than glutaredoxin, as shown for its ortholog from Arabidopsis. The deduced amino acid sequence showed a high degree of similarity to Trx h isoforms from other sources. Phylogenetic studies indicated that VvTrx h2 gene is related to h-type Trx subgroup I. Semi-quantitative RT-PCR analysis revealed that the VvTrx h2 gene was expressed in all plant tissues at different developmental stages.