CRM1 is responsible for intracellular transport mediated by the nuclear export signal

Fukuda, Makoto; Asano, Shiro; Nakamura, Takahiro; Adachi, Makoto; Yoshida, Minoru; Yanagida, Mitsuhiro; Nishida, Eisuke

doi:10.1038/36894

Cited by 1,129 publications

(607 citation statements)

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“…Rev's leucine-rich NES peptide ("LPPLERLTL") was one of the first transferable nuclear export signals to be identified (27)(28)(29) and subsequently played a seminal role in defining CRM1 as the core cellular receptor for NES-bearing cargoes (30)(31)(32). Outside the context of HIV-1 infection, CRM1 regulates the proper nucleocytoplasmic distribution of more than 200 cellular NES-encoding proteins that regulate diverse signaling pathways (30)(31)(32)(33)(34)(35)(36).…”

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confidence: 99%

“…Outside the context of HIV-1 infection, CRM1 regulates the proper nucleocytoplasmic distribution of more than 200 cellular NES-encoding proteins that regulate diverse signaling pathways (30)(31)(32)(33)(34)(35)(36). Moreover, in addition to viral mRNAs, CRM1 regulates the nuclear export of several cellular RNAs, including 5S rRNAs, U snRNAs, and a subset of tightly regulated cellular mRNAs through the activity of NES-bearing adaptor proteins (37).…”

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confidence: 99%

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Cooperativity among Rev-Associated Nuclear Export Signals Regulates HIV-1 Gene Expression and Is a Determinant of Virus Species Tropism

Aligeti

Behrens

Pocock

et al. 2014

J Virol

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Murine cells exhibit a profound block to HIV-1 virion production that was recently mapped to a species-specific structural attribute of the murine version of the chromosomal region maintenance 1 (mCRM1) nuclear export receptor and rescued by the expression of human CRM1 (hCRM1). In human cells, the HIV-1 Rev protein recruits hCRM1 to intron-containing viral mRNAs encoding the Rev response element (RRE), thereby facilitating viral late gene expression. Here we exploited murine 3T3 fibroblasts as a gain-of-function system to study hCRM1's species-specific role in regulating Rev's effector functions. We show that Rev is rapidly exported from the nucleus by mCRM1 despite only weak contributions to HIV-1's posttranscriptional stages. Indeed, Rev preferentially accumulates in the cytoplasm of murine 3T3 cells with or without hCRM1 expression, in contrast to human HeLa cells, where Rev exhibits striking en masse transitions between the nuclear and cytoplasmic compartments. Efforts to bias Rev's trafficking either into or out of the nucleus revealed that Rev encoding a second CRM1 binding domain (Rev-2xNES) or Rev-dependent viral gag-pol mRNAs bearing tandem RREs (GP-2xRRE), rescue virus particle production in murine cells even in the absence of hCRM1. Combined, these results suggest a model wherein Rev-associated nuclear export signals cooperate to regulate the number or quality of CRM1's interactions with viral Rev/RRE ribonucleoprotein complexes in the nucleus. This mechanism regulates CRM1-dependent viral gene expression and is a determinant of HIV-1's capacity to produce virions in nonhuman cell types. IMPORTANCECells derived from mice and other nonhuman species exhibit profound blocks to HIV-1 replication. Here we elucidate a block to HIV-1 gene expression attributable to the murine version of the CRM1 (mCRM1) nuclear export receptor. In human cells, hCRM1 regulates the nuclear export of viral intron-containing mRNAs through the activity of the viral Rev adapter protein that forms a multimeric complex on these mRNAs prior to recruiting hCRM1. We demonstrate that Rev-dependent gene expression is poor in murine cells despite the finding that, surprisingly, the bulk of Rev interacts efficiently with mCRM1 and is rapidly exported from the nucleus. Instead, we map the mCRM1 defect to the apparent inability of this factor to engage Rev multimers in the context of large viral Rev/RNA ribonucleoprotein complexes. These findings shed new light on HIV-1 gene regulation and could inform the development of novel antiviral strategies that target viral gene expression. T he nuclear pore complex (NPC) represents a key barrier to the replication of many viruses that infect eukaryotic cells (1-3). For retroviruses such as the primate lentivirus human immunodeficiency virus type 1 (HIV-1), the late, productive phases of the viral life cycle require efficient nuclear export of unspliced, and thus intron-containing, viral genomic RNAs (gRNAs) that (i) encode the group-specific antigen (Gag) and Gag-Pol polyproteins that form ...

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confidence: 99%

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confidence: 99%

Cooperativity among Rev-Associated Nuclear Export Signals Regulates HIV-1 Gene Expression and Is a Determinant of Virus Species Tropism

Aligeti

Behrens

Pocock

et al. 2014

J Virol

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“…This finding implied that Crm1 functions in the process of protein export from nucleus. Crm1 was subsequently identified as a nuclear export receptor that recognizes the nuclear export signal (NES) sequence in the cargo protein, and LMB as an inhibitor of Crm1-NES binding [46][47][48][49] (Fig. 11-3).…”

Section: Chemical Genetics and Genomics Using Fission Yeastmentioning

confidence: 99%

Chemical Genomics Based on Yeast Genetics

Nishimura

Yashiroda

Yoshida

2009

Protein Targeting With Small Molecules

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“…The export of mRNA and U snRNAs from Xenopus oocyte nuclei appears to require GTP hydrolysis by Ran while the transport of proteins, including HIV-1 Rev and tRNA, is dependent only on the presence of Ran-GTP but not GTP hydrolysis [ 18]. A cellular transport factor, Exportin 1 (previously known as CRM1), has been shown to interact with NES [16], and Exportin 1 mutants in yeast are associated with inhibition of NES-mediated export [32]. Additionally, cooperative binding between the Rev NES, Exportin 1, and RanGTPase has been demonstrated [ 14].…”

Section: Introductionmentioning

confidence: 99%

HIV-1 rev promotes the nuclear export of unspliced and singly spliced RNAs in a mammalian cell-free export system

1999

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Rev has been shown to promote the export of HIV-1 RNAs from Xenopus oocyte nuclei, but a system to examine the direct effect of Rev on HIV-1 RNA export in mammalian somatic cells does not exist. In this report, the development of a cell-free RNA export system using COS cells is described. This system is capable of examining the movement of RNA from nuclei of COS cells transfected with an HIV-1 proviral construct into reconstituted cytosol from nontransfected cells. A reproducible preparation of nuclei free of residual cytoplasmic RNA is demonstrated. Export of RNA from these nuclei into reconstituted cell-free extracts was saturable and dependent on temperature and energy. Further validation of the system was obtained by confirming that the nuclear export of HIV-1-unspliced and partially spliced RNAs was dependent upon the expression of HIV-1 Rev and that the presence of Rev appeared to decrease the export of an HIV-1-spliced RNA. The system was also able to demonstrate that Rev did not appear to significantly enhance the export of an HIV-1 protease-containing RNA that has been shown to be dependent upon Rev for maximal expression. Consequently, the system appears useful for the examination of parameters of nuclear export of HIV-1 and cellular RNAs.

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CRM1 is responsible for intracellular transport mediated by the nuclear export signal

Cited by 1,129 publications

References 28 publications

Cooperativity among Rev-Associated Nuclear Export Signals Regulates HIV-1 Gene Expression and Is a Determinant of Virus Species Tropism

Cooperativity among Rev-Associated Nuclear Export Signals Regulates HIV-1 Gene Expression and Is a Determinant of Virus Species Tropism

Chemical Genomics Based on Yeast Genetics

HIV-1 rev promotes the nuclear export of unspliced and singly spliced RNAs in a mammalian cell-free export system

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