A Conditional Yeast E1 Mutant Blocks the Ubiquitin–Proteasome Pathway and Reveals a Role for Ubiquitin Conjugates in Targeting Rad23 to the Proteasome

E1 ubiquitin activating enzyme catalyzes the initial step in all ubiquitin-dependent processes. We report the isolation of uba1-204, a temperature-sensitive allele of the essential Saccharomyces cerevisiae E1 gene, UBA1. Uba1-204 cells exhibit dramatic inhibition of the ubiquitin–proteasome system, resulting in rapid depletion of cellular ubiquitin conjugates and stabilization of multiple substrates. We have employed the tight phenotype of this mutant to investigate the role ubiquitin conjugates play in the dynamic interaction of the UbL/UBA adaptor proteins Rad23 and Dsk2 with the proteasome. Although proteasomes purified from mutant cells are intact and proteolytically active, they are depleted of ubiquitin conjugates, Rad23, and Dsk2. Binding of Rad23 to these proteasomes in vitro is enhanced by addition of either free or substrate-linked ubiquitin chains. Moreover, association of Rad23 with proteasomes in mutant and wild-type cells is improved upon stabilizing ubiquitin conjugates with proteasome inhibitor. We propose that recognition of polyubiquitin chains by Rad23 promotes its shuttling to the proteasome in vivo

Determination of the platinum – group elements (PGE) and gold (Au) in manganese nodule reference samples by nickel sulfide fire-assay and Te coprecipitation with ICP-MS

7-16Platinum group elements (PGE) and Au data in polymetallic oceanic ferromanganese nodule reference samples and crust samples obtained by inductively coupled plasma mass spectrometry (ICP-MS), after separation and pre-concentration by nickel sulfide fire-assay and Te coprecipitation, are presented. By optimizing several critical parameters such as flux composition, matrix matching calibration, etc., best experimental conditions were established to develop a method suitable for routine analysis of manganese nodule samples for PGE and Au. Calibrations were performed using international PGE reference materials, WMG-1 and WMS-1. This improved procedure offers extremely low detection limits in the range of 0.004 to 0.016 ng/g. The results obtained in this study for the reference materials compare well with previously published data wherever available. New PGE data are also provided on some international manganese nodule reference materials. The analytical methodology described here can be used for the routine analysis of manganese nodule and crust samples in marine geochemical studies

Distribution of cadmium in a cultivated soil in Britanny, France Distribuição de cádmio em solo cultivado na Bretanha, França

Cadmium (Cd) can be potentially toxic to the environment, and its bioavailability is related to the chemical forms it occurs in the soils. The distribution of Cd into the solid phase and its availability was investigated in a cultivated soil in Britanny, France. Cd sequential extraction was performed using a modified Tessier's sequential extraction protocol. Total content of Cd (n = 22 samples) ranged from 0.13 to 0.37 mg kg¹. Long term history of organic and mineral fertilizers application increased Cd concentration in the surface horizon. The fate of Cd was correlated with soil available P2O5 concentration and pH. The regression analysis (linear and non-linear) and Principal Component Analysis revealed the synergistic effect of P2O5 in the retention of Cd in cultivated soils. The following fractions' sequence was observed: bounded to Fe and Al oxides + phosphates > exchangeable > bound to organic matter » residual fraction.<br>O cádmio (Cd) pode ser tóxico no ambiente e sua biodisponibilidade está relacionada às formas químicas em que os metais se encontram no solo. Apresenta-se a distribuição deste metal na fase sólida de um solo cultivado na Bretanha, França, assim como fatores que influenciam sua disponibilidade. Foi realizada a extração sequencial do Cd utilizando o método Tessier, modificado. A concentração total de Cd, avaliada em 22 amostras, variou de 0,13 a 0,37 mg kg¹. O uso contínuo de fertilizantes orgânicos e minerais aumentou a concentração de Cd no horizonte superficial do solo. A distribuição do Cd foi relacionada à concentração de P2O5 disponível e ao pH. Análises de regressão (linear e não linear) e a Análise de Componentes Principais mostraram efeito sinérgico do P2O5 na retenção de Cd nos solos cultivados. O fracionamento apresentou a seguinte sequência: ligado a óxidos de Fe e de Al + fosfatos > trocável > ligado à matéria orgânica » residual