AepG is a glucuronosyltransferase involved in acidic exopolysaccharide synthesis and contributes to environmental adaptation of Haloarcula hispanica.

The attachment of a sugar to a hydrophobic lipid carrier is the first step in the biosynthesis of many glycoconjugates. In the halophilic archaeon Haloarcula hispanica, HAH_1206, renamed AepG, is a predicted glycosyltransferase belonging to the CAZy Group 2 family that shares a conserved amino acid sequence with dolichol phosphate mannose synthases. In this study, the function of AepG was investigated by genetic and biochemical approaches. We found that aepG deletion led to the disappearance of dolichol phosphate-glucuronic acid. Our biochemical assays revealed that recombinant cellulose-binding, domain-tagged AepG could catalyze the formation of dolichol phosphate-glucuronic acid in time- and dose-dependent manners. Based on the in vivo and in vitro analyses, AepG was confirmed to be a dolichol phosphate glucuronosyltransferase involved in the synthesis of the acidic exopolysaccharide produced by H. hispanica. Furthermore, lack of aepG resulted in hindered growth and cell aggregation in high salt medium, indicating that AepG is vital for the adaptation of H. hispanica to a high salt environment. In conclusion, AepG is the first dolichol phosphate glucuronosyltransferase identified in any of the three domains of life and, moreover, offers a starting point for further investigation into the diverse pathways used for extracellular polysaccharide biosynthesis in archaea.

Emp47 and Vip36 are required for polarized growth and protein trafficking between ER and Golgi apparatus in opportunistic fungal pathogen Aspergillus fumigatus.

In Aspergillus fumigatus, an opportunistic fungal pathogen causing fatal invasive aspergillosis, N-glycosylation is vital for polarized growth. To investigate its mechanism, two putative L-type lectin genes emp47 (AFUB_032470) and vip36 (AFUB_027870) were identified in A. fumigatus. Deletion of the emp47 or vip36 gene resulted in delayed germination and abnormal polarity. Also, the Δemp47 displayed an increased resistance to azoles whereas the Δvip36 showed an increased susceptibility to amphotericin B. Secretome analysis revealed that 205 proteins were differentially secreted in the Δemp47 and 145 of them were reduced, while 153 proteins displayed a differential secretion and 134 of them were increased in the Δvip36 as compared with that of the wild-type strain. Also, potential cargo glycoproteins of Emp47 and Vip36 were identified by comparative secretome analysis. Our results suggest that Emp47 is responsible for the transport of proteins from endoplasmic reticulum (ER) to Golgi, while Vip36 acts in protein retrieval from Golgi to ER.

Molecular mechanism of damage and repair of mouse thymus lymphocytes induced by radiation.

OBJECTIVE: To investigate the role of apoptosis in radiation-induced mouse thymus lymphocyte damage and repair and provide the basis for understanding the molecular mechanism of radiation-induced lymphocyte damage and repair as well as the prevention and treatment of acute radiation sickness. METHODS: We studied the dynamic changes of apoptosis of mouse thymus lymphocytes and the expression of bax and bcl-2 gene products after 2, 4, 6 and 8 Gy of whole body gamma-irradiation using in situ terminal labeling, DNA electrophoresis and immunohistochemical techniques. RESULTS: At the early stage after irradiation, the percentage of apoptotic lymphocytes increased rapidly in accordance with the increasing of radiation doses, while the counts of the thymus and peripheral lymphocytes decreased sharply, showing an opposite change to lymphocyte apoptosis. After 6 Gy gamma-irradiation, typical morphological characteristics of thymus apoptotic lymphocytes in early, middle and late stages were found by transmission electron microscopy. The thymus lymphocytes displayed characteristic DNA ladders 4 hr and 8 hr after 2-6 Gy gamma-irradiation,using DNA gel electrophoresis techniques. Abnormal expression of bcl-2 and bax gene products were shown in irradiated lymphocytes. CONCLUSIONS: Apoptosis plays an important role in the process of radiation-induced mouse thymus lymphocyte damage and repair. Bcl-2 and Bax proteins may regulate the process of lymphocyte apoptosis.