Hepatitis C virus triggers Golgi fragmentation and autophagy through the immunity-related GTPase M.

Positive-stranded RNA viruses, such as hepatitis C virus (HCV), assemble their viral replication complexes by remodeling host intracellular membranes to a membranous web. The precise composition of these replication complexes and the detailed mechanisms by which they are formed are incompletely understood. Here we show that the human immunity-related GTPase M (IRGM), known to contribute to autophagy, plays a previously unrecognized role in this process. We show that IRGM is localized at the Golgi apparatus and regulates the fragmentation of Golgi membranes in response to HCV infection, leading to colocalization of Golgi vesicles with replicating HCV. Our results show that IRGM controls phosphorylation of GBF1, a guanine nucleotide exchange factor for Arf-GTPases, which normally operates in Golgi membrane dynamics and vesicle coating in resting cells. We also find that HCV triggers IRGM-mediated phosphorylation of the early autophagy initiator ULK1, thereby providing mechanistic insight into the role of IRGM in HCV-mediated autophagy. Collectively, our results identify IRGM as a key Golgi-situated regulator that links intracellular membrane remodeling by autophagy and Golgi fragmentation with viral replication.

Effects on development, growth responses and thyroid-hormone systems in eyed-eggs and yolk-sac larvae of Atlantic salmon (Salmo salar) continuously exposed to 3,3',4,4'-tetrachlorobiphenyl (PCB-77).

Thyroid hormones (triiodothyronine, T3; and thyroxine, T4) play significant roles in development, metamorphosis, metabolism, homeostasis, cellular proliferation, and differentiation, for which the effects are mediated through thyroid hormone receptors (TRα and TRß). Similarly, the insulin-like growth factor (IGF) is involved in growth and development through regulation of somatic growth. This study was designed to examine the effects of the dioxin-like 3,3',4,4'-tetrachlorobiphenyl (PCB-77) on responses related to growth and thyroid hormone system in eyed eggs and yolk-sac larvae of Atlantic salmon. Salmon eggs were continuously exposed to two waterborne concentrations of PCB-77 (1 or 10 ng/L) over a period of 50 d covering hatching and through yolk-sac absorption stages. Sampling was performed regularly throughout the exposure period and at different time intervals. Gene expression patterns were performed on whole-body homogenate at age 500, 548, 632, 674, and 716 dd (dd: day degrees) using quantitative polymerase chain reaction (PCR). Total T3 (TT3) and total T4 (TT4) were measured using radioimmunoassay (RIA). Data showed that 10 ng PCB-77 increased dioiodinase 2 (Dio2) at 500 dd and both PCB-77 concentrations decreased dio2 expression at 548 dd. PCB-77 elevated cellular TT3 at 500 dd and was lowered at 548 dd only at 10 ng. Otherwise, time-related reduction was not affected by PCB-77 exposure as observed for the rest of the exposure period. For TT4, 1 ng PCB-77 produced a rise at 500 dd, and an apparent concentration decrease at 548 dd, before a total inhibition at 632 dd. The IGF-1 and IGF-1R were variably affected by PCB-77. For IGF-2, PCB-77 produced a concentration-dependent increase at 548 dd, and thereafter an elevation (1 ng) and fall (10 ng) at 632 dd. TRß mRNA demonstrated PCB-77 related increases during the exposure period, and this effect returned to control levels at 716 dd. For TRα, a rise was noted only after exposure to 10 ng PCB-77 at 500 dd. Overall, the present study demonstrates some possible growth and developmental consequences following exposure to PCB-77 during early life stages of Atlantic salmon.

Neural aromatase transcript and protein levels in Atlantic salmon (Salmo salar) are modulated by the ubiquitous water pollutant, 4-nonylphenol.

At present, there are no known direct occurrences of nonylphenol (NP) in nature. Therefore, its presence in nature is solely a consequence of human activities. NP is generated through degradation of alkylphenol ethoxylates released mainly from textile, metal working, institutional cleansing and laundry cleaning, but few data on the amount of the release is available. These compounds have been shown to affect several biological processes, including the endocrine systems, in a wide number of species. The cytochrome P450 aromatase (Cyp19) is the rate-limiting step in estrogen production, and is known to be a potential target for endocrine-disrupting chemicals (EDCs) such as NP. Teleost fish generally have a high brain aromatase activity, and the effects of EDCs in fish brain is not thoroughly investigated. In this study, juvenile Atlantic salmon (Salmo salar) were exposed to waterborne concentrations of the synthetic pharmaceutical and xenoestrogen 17alpha-ethynylestradiol (EE2; 5ng/L) and the xenoestrogen 4-nonylphenol (NP; 5 and 50microg/L) for 72h. Brain tissue and blood were sampled from individual fish. Gene expression patterns of Cyp19 isoforms were determined by quantitative PCR, aromatase protein immunoreactivity in the brain was evaluated by immunohistochemistry and immunoblotting, and aromatase activity was analyzed using the tritiated water-release assay. Plasma estradiol (E2) and testosterone (T) levels were measured by EIA. In the brain, EE2 increased the mRNA expression of Cyp19b almost threefold compared to the solvent control, whereas Cyp19a levels were unaffected by EE2 treatment. In contrast, both NP concentrations produced significant reduction of Cyp19a expression. Immunohistochemical aromatase protein reactivity was localized in several brain regions, but no apparent quantitative effects of the exposures were observed. Immunoblotting analysis showed that EE2 and NP produced a slight increase in brain immunoreactive aromatase protein band, compared with controls. Plasma levels of E2 increased twofold when treated with EE2 and 5microg NP/L, and threefold when exposed to 50microg NP/L. In general, the present study shows that the parallel biochemical, transcriptional and cellular detection of neural aromatase for endocrine-disrupting effects from EE2 and NP may be observed at specific levels of the biological organization.