␤-Catenin is a member of the Armadillo repeat protein family with a dual cellular function as a component of both the adherens junction complex and the Wnt/wingless signaling pathway. Here we show that ␤-catenin is proteolytically cleaved during anoikis and staurosporine-induced apoptosis. Cleavage of ␤-catenin was found to be caspase-dependent. Five cleavage products of ␤-catenin were identified in vivo and after in vitro cleavage by caspase-3. Amino acid sequencing and mass spectrometry analysis indicated two caspase-3 cleavage sites at the C terminus and three further sites at the N terminus, whereas the central Armadillo repeat region remained unaffected. All ␤-catenin cleavage products were still able to associate with E-cadherin and ␣-catenin and were found to be enriched in the cytoplasm. Functional analysis revealed that ␤-catenin deletion constructs resembling the observed proteolytic fragments show a strongly reduced transcription activation potential when analyzed in gene reporter assays. We therefore conclude that an important role of the ␤-catenin cleavage during apoptosis is the removal of its transcription activation domains to prevent its transcription activation potential.

The amino acid composition of the coat proteins of the following viruses is reported: Andean potato latent, clitoria yellow vein, desmodium yellow mottle, dulcamara mottle, eggplant mosaic, okra mosaic, ononis yellow mosaic, scrophularia mottle, and, as controls, cocksfoot mild mosaic and cocksfoot mottle viruses. These data, together with some already published, were used to compute classifications of the tymoviruses. These classifications show a general similarity to Koenig’s serological classification of the tymoviruses, but the correlation is poor, unlike similar comparisons of tobamovirus classifications. Several possible reasons for the poor correlation have been examined and excluded, and its implications are discussed.

Dalcochinin-8'-O-beta-glucoside beta-glucosidase (dalcochinase) from the Thai rosewood (Dalbergia cochinchinensis Pierre) has aglycone specificity for isoflavonoids and can hydrolyze both beta-glucosides and beta-fucosides. To determine its structure and evolutionary lineage, the sequence of the enzyme was determined by peptide sequencing followed by PCR cloning. The cDNA included a reading frame coding for 547 amino acids including a 23 amino acid propeptide and a 524 amino acid mature protein. The sequences determined at peptide level were found in the cDNA sequence, indicating the sequence obtained was indeed the dalcochinase enzyme. The mature enzyme is 60% identical to the cyanogenic beta-glucosidase from white clover glycosyl hydrolase family 1, for which an X-ray crystal structure has been solved. Based on this homology, residues which may contribute to the different substrate specificities of the two enzymes were identified. Eight putative glycosylation sites were identified, and one was confirmed to be glycosylated by Edman degradation and mass spectrometry. The protein was expressed as a prepro-alpha-mating factor fusion in Pichia pastoris, and the activity of the secreted enzyme was characterized. The recombinant enzyme and the enzyme purified from seeds showed the same K(m) for pNP-glucoside and pNP-fucoside, had the same ratio of V(max) for these substrates, and similarly hydrolyzed the natural substrate, dalcochinin-8'-beta-glucoside.

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