Biochemical Properties and Biological Functions of FET Proteins

Schwartz, Jacob C.; Cech, Thomas R.; Parker, Roy

doi:10.1146/annurev-biochem-060614-034325

Cited by 193 publications

(257 citation statements)

References 147 publications

(354 reference statements)

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“…Moreover, we recently obtained evidence that the U1 snRNP core complex (U1 snRNA and the Sm proteins) is comislocalized to the cytoplasm with FUS in ALS patient fibroblasts harboring mutations in the NLS (40). Thus, decreased levels of both FUS and U1 snRNP in the nucleus may lead to the aberrant splicing that has been reported in transfected cells and ALS patient cells expressing FUS with NLS mutations (29,30,38,39). In previous work, FUS was shown to bind to the C-terminal domain of RNAP II and regulate phosphorylation of serine-2 (23,25,27).…”

Section: Fus Is Required For the Interaction Between Rnap II And U1 Smentioning

confidence: 87%

“…Considering that transcription-coupled splicing is such a fundamental cellular process, it is possible that its disruption by mutant FUS contributes to the pathogenesis of ALS. In support of this possibility, many of the mutations in FUS that are ALScausative are located in the nuclear localization signal (NLS), which leads to mislocalization of FUS to the cytoplasm in ALS patient cells (29,38,39). Moreover, we recently obtained evidence that the U1 snRNP core complex (U1 snRNA and the Sm proteins) is comislocalized to the cytoplasm with FUS in ALS patient fibroblasts harboring mutations in the NLS (40).…”

Section: Fus Is Required For the Interaction Between Rnap II And U1 Smentioning

confidence: 95%

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FUS functions in coupling transcription to splicing by mediating an interaction between RNAP II and U1 snRNP

Reed

2015

Proc. Natl. Acad. Sci. U.S.A.

100

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Pre-mRNA splicing is coupled to transcription by RNA polymerase II (RNAP II). We previously showed that U1 small nuclear ribonucleoprotein (snRNP) associates with RNAP II, and both RNAP II and U1 snRNP are also the most abundant factors associated with the protein fused-in-sarcoma (FUS), which is mutated to cause the neurodegenerative disease amyotrophic lateral sclerosis. Here, we show that an antisense morpholino that base-pairs to the 5′ end of U1 snRNA blocks splicing in the coupled system and completely disrupts the association between U1 snRNP and both FUS and RNAP II, but has no effect on the association between FUS and RNAP II. Conversely, we found that U1 snRNP does not interact with RNAP II in FUS knockdown extracts. Moreover, using these extracts, we found that FUS must be present during the transcription reaction in order for splicing to occur. Together, our data lead to a model that FUS functions in coupling transcription to splicing via mediating an interaction between RNAP II and U1 snRNP.ALS | coupling transcription to splicing | RNA polymerase II | U1 snRNP I t is now well established that the steps in gene expression are extensively coupled to one another, including both physical and functional coupling between RNA polymerase II (RNAP II) transcription and pre-mRNA processing (1-11). Moreover, the majority of nascent transcripts are spliced cotranscriptionally, while the transcripts are still tethered to RNAP II. In vitro systems have been developed for the coupled transcription/splicing (txn/splicing) reaction as well as for cotranscriptional splicing, and these systems have been used to investigate the mechanisms underlying these processes (12-17). In the case of coupled txn/ splicing, studies using the in vitro system revealed that transcription potently enhances spliceosome assembly, which in turn leads to a strong enhancement of the splicing reaction (13). Additional studies revealed that the only essential splicing factors that copurify with RNAP II are U1 small nuclear ribonucleoprotein (snRNP) and its associated factors, the serine/ arginine-rich (SR) proteins (18)(19)(20). This observation is particularly noteworthy because U1 snRNP/SR proteins are the first splicing factors that bind to pre-mRNA during spliceosome assembly (21). Although functional studies indicate that SR proteins play a key role in coupling transcription to splicing (19), the role of U1 snRNP in this coupling event has not been examined. It is also not known how U1 snRNP interacts with RNAP II. U1 snRNA is known to base-pair to the 5′ splice site during the earliest steps in spliceosome assembly, and this interaction is essential for splice-site recognition (21). In addition, we and others found that U1 snRNP/SR and RNAP II are among the main factors that associate with the protein fused-in-sarcoma (FUS) (19,(22)(23)(24)(25)(26)(27). Understanding the normal roles of FUS and the pathways in which it functions are of great importance because FUS is mutated to cause the fatal motor neuron disease amyotrophic lateral s...

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Section: Fus Is Required For the Interaction Between Rnap II And U1 Smentioning

confidence: 87%

Section: Fus Is Required For the Interaction Between Rnap II And U1 Smentioning

confidence: 95%

FUS functions in coupling transcription to splicing by mediating an interaction between RNAP II and U1 snRNP

Reed

2015

Proc. Natl. Acad. Sci. U.S.A.

100

View full text Add to dashboard Cite

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“…As described above, RRM, ZnF, and RGG motifs in FET proteins play major roles in the binding of RNA (Schwartz et al 2015). While the RRM alone exhibited very weak affinity to RNA, inclusion of the flanking RGG motifs significantly strengthened the RNA binding of RRM (Liu et al 2013;Schwartz et al 2013;Lerga et al 2001).…”

Section: Introductionmentioning

confidence: 84%

“…(Riggi et al 2007). Also, the FET proteins have the same domain organization: an N-terminal region with low amino acid complexity that is rich in Gln, Gly, Ser, and Tyr (LC domain), an RNA recognition motif (RRM), a Zn-finger motif (ZnF), three regions rich in Arg and Gly (RGG1, 2, and 3), and a nuclear localization signal called PY-NLS at the C-terminus (Schwartz et al 2015). Many ALS-causing mutations are reported in the PY-NLS of FUS (Mackenzie et al 2010a), which retards the nuclear transporting function and thus results in the abnormal accumulation of mutant FUS in the cytoplasm.…”

Section: Introductionmentioning

confidence: 99%

“…FET proteins can bind single-stranded DNA (ssDNA), double-stranded DNA (dsDNA), and RNA. FUS has been reported to bind ssDNA more tightly than dsDNA in vitro (Schwartz et al 2015) and shows the ability to anneal DNA strands and promote ssDNA invasion of dsDNA (Bertrand et al 1999;Baechtold et al 1999). The apparent annealing activity of FET proteins may thus be involved in the DNA break repair; indeed, a knockout of the FUS gene in mice has been shown to accumulate DNA breaks after treatment with ionizing radiation (Kuroda et al 2000).…”

Section: Introductionmentioning

confidence: 99%

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Aggregation of FET Proteins as a Pathological Change in Amyotrophic Lateral Sclerosis

Furukawa

Tokuda

2016

Advances in Experimental Medicine and Biology

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Amyotrophic lateral sclerosis (ALS) is a fatal motor neuron disease that is characterized by the formation of abnormal inclusions in neurons. While the pathomechanism of ALS remains obscure, a number of proteins have been identified in the inclusion bodies, and the pathological roles of RNA-binding proteins have been increasingly emphasized. Among those, the FET proteins (FUS, EWSR1, TAF15) were recently identified as RNA-binding proteins in pathological inclusions of ALS and other neurodegenerative diseases; moreover, mutations in the genes encoding the FET proteins were found to be associated with familial forms of ALS. FET proteins are normally localized in the nucleus, but the introduction of pathogenic mutations in FET proteins leads to their abnormal redistribution to the cytoplasm, where they form aggregates. While further investigation will be required to understand the intracellular factors controlling the aggregation propensities of FET proteins, they are thought to lose their physiological functions and become toxic through their misfolding/aggregation. Here, we will briefly review recent advances of our understanding of the physiological functions and aggregation behavior of FET proteins in vivo as well as in vitro.

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FET fusion oncoproteins interact with BRD4 and SWI/SNF chromatin remodelling complex subtypes in sarcoma

et al. 2022

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FET fusion oncoproteins containing one of the FET (FUS, EWSR1, TAF15) family proteins juxtaposed to alternative transcription-factor partners are characteristic of more than 20 types of sarcoma and leukaemia. FET oncoproteins bind to the SWI/SNF chromatin remodelling complex, which exists in three subtypes: cBAF, PBAF and GBAF/ncBAF. We used comprehensive biochemical analysis to characterize the interactions between FET oncoproteins, SWI/SNF complexes and the transcriptional coactivator BRD4. Here, we report that FET oncoproteins bind all three main SWI/SNF subtypes cBAF, PBAF and GBAF, and that FET oncoproteins interact indirectly with BRD4 via their shared interaction partner SWI/SNF. Furthermore, chromatin immunoprecipitation sequencing and proteomic analysis showed that FET oncoproteins, SWI/SNF components and BRD4 co-localize on chromatin and interact with mediator and RNA Polymerase II. Our results provide a possible molecular mechanism for the FET-fusion-induced oncogenic transcriptional profiles and may lead to novel therapies targeting aberrant SWI/SNF complexes and/or BRD4 in FET-fusion-caused malignancies.

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Biochemical Properties and Biological Functions of FET Proteins

Cited by 193 publications

References 147 publications

FUS functions in coupling transcription to splicing by mediating an interaction between RNAP II and U1 snRNP

FUS functions in coupling transcription to splicing by mediating an interaction between RNAP II and U1 snRNP

Aggregation of FET Proteins as a Pathological Change in Amyotrophic Lateral Sclerosis

FET fusion oncoproteins interact with BRD4 and SWI/SNF chromatin remodelling complex subtypes in sarcoma

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