HylA, an alternative hydrolase for initiation of catabolism of the phenylurea herbicide linuron in Variovorax sp. strains.

Variovorax sp. strain WDL1, which mineralizes the phenylurea herbicide linuron, expresses a novel linuron-hydrolyzing enzyme, HylA, that converts linuron to 3,4-dichloroaniline (DCA). The enzyme is distinct from the linuron hydrolase LibA enzyme recently identified in other linuron-mineralizing Variovorax strains and from phenylurea-hydrolyzing enzymes (PuhA, PuhB) found in Gram-positive bacteria. The dimeric enzyme belongs to a separate family of hydrolases and differs in Km, temperature optimum, and phenylurea herbicide substrate range. Within the metal-dependent amidohydrolase superfamily, HylA and PuhA/PuhB belong to two distinct protein families, while LibA is a member of the unrelated amidase signature family. The hylA gene was identified in a draft genome sequence of strain WDL1. The involvement of hylA in linuron degradation by strain WDL1 is inferred from its absence in spontaneous WDL1 mutants defective in linuron hydrolysis and its presence in linuron-degrading Variovorax strains that lack libA. In strain WDL1, the hylA gene is combined with catabolic gene modules encoding the downstream pathways for DCA degradation, which are very similar to those present in Variovorax sp. SRS16, which contains libA. Our results show that the expansion of a DCA catabolic pathway toward linuron degradation in Variovorax can involve different but isofunctional linuron hydrolysis genes encoding proteins that belong to evolutionary unrelated hydrolase families. This may be explained by divergent evolution and the independent acquisition of the corresponding genetic modules.

Tumour cellularity, oestrogen receptors and prognosis in breast cancer.

One hundred and thirty-seven patients with operable breast cancer have been treated by mastectomy and node sampling. At the time of operation, portions of tumour were taken for determination of oestrogen receptor activity. In 91 cases adjacent tissue was taken for fixation and histological assessment of malignant epithelial cell content ('cellularity index'). The patients were followed for two years or more, or until first recurrence or death. Prognosis (probability of staying disease-free) was significantly better in patients whose tumours contained receptors than in those whose tumours were receptor-negative (p less than 0.03). However, the best prognosis was in patients with low receptor positive tumours. Those with highest values had a prognosis almost as poor as patients with receptor-negative tumours. In addition, a new finding emerged: prognosis was better in patients with tumours of low cellularity than in those with tumours of high cellularity (p less than 0.04). Since, for receptor-positive tumours, receptor level was also significantly correlated with cellularity (R = +0.35, p less than 0.01), the relationship between prognosis and receptor level was re-examined after 'correction' of the latter quantity for differences in cellularity: on this basis, again prognosis was better in patients with receptor-positive tumours (p less than 0.02), but there was no longer a tendency for any one sub-group of patients with receptor-positive tumours to fare better than the rest. It is concluded that prognosis in operable breast cancer is related to oestrogen receptor activity (either per mg weight, or per cell) and to cellularity.