Tyrosine Phosphorylation in the C-Terminal Nuclear Localization and Retention Signal (C-NLS) of the EWS Protein

Leemann-Zakaryan, Ruzanna P.; Pahlich, Steffen; Grossenbacher, Doris; Gehring, Heinz

doi:10.1155/2011/218483

Cited by 26 publications

(19 citation statements)

References 19 publications

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“…Once in the cytosol, FET proteins are released from their bound mRNAs by one of the following: mRNP remodeling components, posttranslational modifications that decrease RNA binding, displacement by elongating ribosomes, or ultimately degradation of the bound mRNA. Cytoplasmic FET proteins are then reimported into the nucleus upon transportin recognition of a nontraditional nuclear localization signal (PY-NLS) found at the C terminus of FET proteins (118)(119)(120). Modifications near this PY-NLS sequence, including arginine methylation and tyrosine phosphorylation, alter the cytoplasmic accumulation of the FET proteins (96-98, 118, 120-123).…”

Section: Nuclear-cytoplasmic Shuttling Of Fet Proteinsmentioning

confidence: 99%

Biochemical Properties and Biological Functions of FET Proteins

Schwartz

Cech

Parker

2015

Annu. Rev. Biochem.

194

240

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Members of the FET protein family, consisting of FUS, EWSR1, and TAF15, bind to RNA and contribute to the control of transcription, RNA processing, and the cytoplasmic fates of messenger RNAs in metazoa. FET proteins can also bind DNA, which may be important in transcription and DNA damage responses. FET proteins are of medical interest because chromosomal rearrangements of their genes promote various sarcomas and because point mutations in FUS or TAF15 can cause neurodegenerative diseases such as amyotrophic lateral sclerosis and frontotemporal lobar dementia. Recent results suggest that both the normal and pathological effects of FET proteins are modulated by low-complexity or prion-like domains, which can form higher-order assemblies with novel interaction properties. Herein, we review FET proteins with an emphasis on how the biochemical properties of FET proteins may relate to their biological functions and to pathogenesis.

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Section: Nuclear-cytoplasmic Shuttling Of Fet Proteinsmentioning

confidence: 99%

Biochemical Properties and Biological Functions of FET Proteins

Schwartz

Cech

Parker

2015

Annu. Rev. Biochem.

194

240

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“…EWSR1 is an RNA-binding protein that shuttles between the nucleus and cytoplasm, but is predominantly nuclear (29)(30)(31). Given that HCV replicates in the cytoplasm, we next examined the localization of EWSR1 in HCV-infected cells.…”

Section: Ewsr1 Binds Hcv Rna Elementsmentioning

confidence: 99%

“…We identified a novel interaction between the cellular RNA-binding protein EWSR1 and the HCV CRE. EWSR1 is a predominantly nuclear protein that shuttles between the nucleus and cytoplasm (29)(30)(31) and has been previously identified as a potential HCV cofactor in a genome-wide siRNA screen (32). It is a member of the TET family, which regu-lates transcription and RNA processing (33,34).…”

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confidence: 99%

EWSR1 Binds the Hepatitis C Virus cis -Acting Replication Element and Is Required for Efficient Viral Replication

Oakland

Haselton

Randall

2013

J Virol

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The hepatitis C virus (HCV) genome contains numerous RNA elements that are required for its replication. Most of the identified RNA structures are located within the 5= and 3= untranslated regions (UTRs). One prominent RNA structure, termed the cis-acting replication element (CRE), is located within the NS5B coding region. Mutation of part of the CRE, the 5BSL3.2 stemloop, impairs HCV RNA replication. This loop has been implicated in a kissing interaction with a complementary stem-loop structure in the 3= UTR. Although it is clear that this interaction is required for viral replication, the function of the interaction, and its regulation are unknown. In order to gain insight into the CRE function, we isolated cellular proteins that preferentially bind the CRE and identified them using mass spectrometry. This approach identified EWSR1 as a CRE-binding protein. Silencing EWSR1 expression impairs HCV replication and infectious virus production but not translation. While EWRS1 is a shuttling protein that is extensively nuclear in hepatocytes, substantial amounts of EWSR1 localize to the cytosol in HCV-infected cells and colocalize with sites of HCV replication. A subset of EWRS1 translocates into detergent-resistant membrane fractions, which contain the viral replicase proteins, in cells with replicating HCV. EWSR1 directly binds the CRE, and this is dependent on the intact CRE structure. Finally, EWSR1 preferentially interacts with the CRE in the absence of the kissing interaction. This study implicates EWSR1 as a novel modulator of CRE function in HCV replication. Hepatitis C virus (HCV) is a major cause of chronic hepatitis, cirrhosis, and hepatocellular carcinoma. HCV is an enveloped, positive-strand RNA virus classified within the family Flaviviridae. The viral genome encodes an open reading frame of ϳ3,011 codons that is translated as a single polyprotein, which is cleaved by viral and host proteases into at least 10 distinct products. The HCV RNA genome is flanked by the 5= and 3= untranslated regions (UTR). An internal ribosome entry site (IRES) within the 5=UTR is essential for translational initiation of the viral RNA (1, 2). Other RNA elements in the 5=UTR, stem-loop 1, and the 3=UTR, including a variable poly-U/UC tract and three stem-loops contained in the terminal 98 nucleotides that are referred to as the 3=X region, are indispensable for RNA replication (3-7). Multiple viral and host proteins have been suggested to interact with the HCV 3=UTR. Components of the viral replicase complex, NS3, NS5A, and NS5B, have each been shown biochemically to interact with the 3=UTR (8-10). Several cellular proteins have also been implicated in 3=UTR binding, including polypyrimidine tract binding protein (PTB), heterogeneous nuclear ribonucleoprotein C, glyceraldehyde dehydrogenase, HuR, and La (11-17). PTB, HuR, and La have also been shown to be required for efficient HCV RNA replication (18-21). These have been proposed to play multiple roles in HCV replication, including the regulation of translation versus the ...

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“…It has been previously reported that FUS undergoes arginine methylation in arginine-glycine-glycine (RGG) domains near to the NLS that impairs TNPO1 binding to RGG domains, inhibiting nuclear import (Rappsilber et al, 2003;Hung et al, 2009;Dormann et al, 2012;Tradewell et al, 2012;Yamaguchi and Kitajo, 2012;Scaramuzzino et al, 2013). Finally, phosphorylation of the C-terminal Y656 residue within the NLS of the EWSR1 protein has been shown to inhibit nuclear import (Leemann-Zakaryan et al, 2011). As FUS and EWSR1 have highly homologous NLS domains, we sought to explore the functional effects of post-translational modifications of the NLS of FUS.…”

Section: Introductionmentioning

confidence: 99%

Phosphorylation of C-terminal tyrosine 526 in FUS impairs its nuclear import

Darovic

Mihevc

Župunski

et al. 2015

Journal of Cell Science

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Aberrant cytoplasmic aggregation of FUS, which is caused by mutations primarily in the C-terminal nuclear localisation signal, is associated with 3% of cases of familial amyotrophic lateral sclerosis (ALS). FUS aggregates are also pathognomonic for 10% of all frontotemporal lobar degeneration (FTLD) cases; however, these cases are not associated with mutations in the gene encoding FUS. This suggests that there are differences in the mechanisms that drive inclusion formation of FUS in ALS and FTLD. Here, we show that the C-terminal tyrosine residue at position 526 of FUS is crucial for normal nuclear import. This tyrosine is subjected to phosphorylation, which reduces interaction with transportin 1 and might consequentially affect the transport of FUS into the nucleus. Furthermore, we show that this phosphorylation can occur through the activity of the Src family of kinases. Our study implicates phosphorylation as an additional mechanism by which nuclear transport of FUS might be regulated and potentially perturbed in ALS and FTLD.

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Tyrosine Phosphorylation in the C-Terminal Nuclear Localization and Retention Signal (C-NLS) of the EWS Protein

Cited by 26 publications

References 19 publications

Biochemical Properties and Biological Functions of FET Proteins

Biochemical Properties and Biological Functions of FET Proteins

EWSR1 Binds the Hepatitis C Virus cis -Acting Replication Element and Is Required for Efficient Viral Replication

Phosphorylation of C-terminal tyrosine 526 in FUS impairs its nuclear import

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