Journal
BIOSCIENCE REPORTS
Volume 32, Issue 2, Pages 171-184Publisher
PORTLAND PRESS LTD
DOI: 10.1042/BSR20110069
Keywords
autophagy; coat protein; endoplasmic reticulum stress; fluorescence in situ hybridization; tobacco mosaic virus (TMV) RNA; transmission electron microscopy
Categories
Funding
- National Natural Science Foundation of China [30730054, 30572119, 30670937, 30971279, 30901363]
- Ministry of Science and Technology [2007AA02Z120]
- Ministry of Education [20060486008, 20090141120011]
- National Innovation Experiment Program for College Students [WU2007061]
- Provincial Department of Science and Technology of Hubei, China [2007ABC010]
- Ministry of Education, China
- Li Ka Shing Foundation, Hong Kong, China
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The ability of human cells to defend against viruses originating from distant species has long been ignored. Owing to the pressure of natural evolution and human exploration, some of these viruses may be able to invade human beings. If their 'fresh' host had no defences, the viruses could cause a serious pandemic, as seen with HIV, SARS (severe acute respiratory syndrome) and avian influenza virus that originated from chimpanzees, the common palm civet and birds, respectively. It is unknown whether the human immune system could tolerate invasion with a plant virus. To model such an alien virus invasion, we chose TMV (tobacco mosaic virus) and used human epithelial carcinoma cells (He La cells) as its 'fresh' host. We established a reliable system for transfecting TMV-RNA into He La cells and found that TMV-RNA triggered autophagy in He La cells as shown by the appearance of autophagic vacuoles, the conversion of LC3-I (light chain protein 3-I) to LC3-II, the up-regulated expression of Beclin1 and the accumulation of TMV protein on autophagosomal membranes. We observed suspected TMV virions in He La cells by TEM (transmission electron microscopy). Furthermore, we found that TMV-RNA was translated into CP (coat protein) in the ER (endoplasmic reticulum) and that TMV-positive RNA translocated from the cytoplasm to the nucleolus. Finally, we detected greatly increased expression of GRP78 (78 kDa glucose-regulated protein), a typical marker of ERS (ER stress) and found that the formation of autophagosomes was closely related to the expanded ER membrane. Taken together, our data indicate that HeLa cells used ERS and ERS-related autophagy to defend against TMV-RNA.
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