Quantitative Proteomics Using Stable Isotope Labeling with Amino Acids in Cell Culture Reveals Changes in the Cytoplasmic, Nuclear, and Nucleolar Proteomes in Vero Cells Infected with the Coronavirus Infectious Bronchitis Virus

Emmott, Edward; Rodgers, Mark; Macdonald, Andrew; McCrory, Sarah; Ajuh, Paul; Hiscox, Julian A.

doi:10.1074/mcp.m900345-mcp200

Cited by 79 publications

(108 citation statements)

References 85 publications

(92 reference statements)

Supporting

Mentioning

103

Contrasting

Order By: Relevance

“…Nevertheless, SERBP1 is not included in the NOPdb (Nucleolar Proteome Database), which includes more than 4500 human proteins identified by mass spectrometry from purified nucleoli [24]. No nucleolar distribution of SERBP1 was suggested by nucleolar proteomic analyzes of the dynamic distribution under virus infection [25,26] or DNA damage [27]. However, two nucleolar localization signals in the N-terminus were determined using NoD [28].…”

Section: Discussionmentioning

confidence: 99%

Localization of SERBP1 in stress granules and nucleoli

et al. 2013

View full text Add to dashboard Cite

SERPINE1 mRNA-binding protein 1 (SERBP1) is an arginine-methylated RNA-binding protein whose modification affects protein interaction and intracellular localization. In the present study, we show that, under normal growth conditions without stress, SERBP1 interacts with arginine-methylated and stress granule-associated proteins such as heterogeneous nuclear ribonucleoprotein A1, fragile X mental retardation protein and fragile X mental retardation syndrome-related protein 1 in an RNA-dependent manner. We also show that, after arsenite treatment, a proportion of full-length SERBP1 protein co-localizes with the typical stress granule marker T-cell intracellular antigen-1 in the cytoplasmic stress granules. Truncated SERBP1 with an Nterminal, central RG or C-terminal deletion, or single-domain segments comprising the N-terminal, central or C-terminal region, were recruited to stress granules upon arsenite treatment but with reduced efficiency. In addition, upon arsenite treatment, the localization of SERBP1 changed from a diffuse cytoplasmic localization to nuclear-dominant (concentrated in the nucleolus) A similar distribution was observed when cells were treated with the methylation inhibitor adenosine periodate, and was also detected for N-or C-terminal domain deletions and all three single-domain fragments even without stress induction. We further demonstrate that adenosine periodate treatment delays the association/dissociation of SERBP1 with stress granules. Hypomethylation retains SERBP1 in the nucleus/nucleolus regardless of arsenite treatment. Our study indicates that arginine methylation is correlated with recruitment of SERBP to stress granules and nucleoli and its retention therein. To our knowledge, this is the first report of an RNA-binding protein that is shifted simultaneously to cytoplasmic stress granules and nucleoli, two ribonucleoprotein-enriched subcellular compartments, upon stress.

show abstract

Section: Discussionmentioning

confidence: 99%

Localization of SERBP1 in stress granules and nucleoli

et al. 2013

View full text Add to dashboard Cite

show abstract

“…However, there is ample evidence that implicates the nucleus and nuclear proteins in the replication and pathogenesis of positive-strand RNA viruses, including coronaviruses (64). The replication of these RNA viruses in enucleated cells is variable, ranging from 10% to 100% of that in nucleated controls (65, 66).…”

Section: Relevance Of the Cell Nucleus In Coronavirus Rna Synthesismentioning

confidence: 99%

“…The replication of these RNA viruses in enucleated cells is variable, ranging from 10% to 100% of that in nucleated controls (65, 66). The relocation of nuclear proteins to the cytoplasm and of viral proteins to the nucleus during virus replication (7, 64, 67) (Table 1) highlights the relevance of this organelle during the coronavirus infectious cycle and raises important questions: What is the role of nuclear factors in the replication of these viruses, and do viral proteins traveling to the nucleus participate in RTC activity?…”

Section: Relevance Of the Cell Nucleus In Coronavirus Rna Synthesismentioning

confidence: 99%

Continuous and Discontinuous RNA Synthesis in Coronaviruses

et al. 2015

View full text Add to dashboard Cite

Replication of the coronavirus genome requires continuous RNA synthesis, whereas transcription is a discontinuous process unique among RNA viruses. Transcription includes a template switch during the synthesis of subgenomic negative-strand RNAs to add a copy of the leader sequence. Coronavirus transcription is regulated by multiple factors, including the extent of base-pairing between transcription-regulating sequences of positive and negative polarity, viral and cell protein–RNA binding, and high-order RNA-RNA interactions. Coronavirus RNA synthesis is performed by a replication-transcription complex that includes viral and cell proteins that recognize cis-acting RNA elements mainly located in the highly structured 5′ and 3′ untranslated regions. In addition to many viral nonstructural proteins, the presence of cell nuclear proteins and the viral nucleocapsid protein increases virus amplification efficacy. Coronavirus RNA synthesis is connected with the formation of double-membrane vesicles and convoluted membranes. Coronaviruses encode proofreading machinery, unique in the RNA virus world, to ensure the maintenance of their large genome size.

show abstract

“…Quantitative proteomic analysis can also be used to profile changes in protein abundance in response to viral infection as well as virus-host protein interactions. 295 Emmott et al identified and quantified 1830 proteins in control and coronavirus infectious bronchitis v irus (IBV)-infected Vero cells utilizing subcellular fractionation and S ILAC labeling. Coronaviruses can cause respiratory as well as gastrointestinal infections in humans and animals.…”

Section: Organelle Compositionmentioning

confidence: 99%