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Volumn 10, Issue 3, 1998, Pages 400-408

Functions of the GTPase Ran in RNA export from the nucleus

Author keywords

[No Author keywords available]

Indexed keywords

GUANOSINE DIPHOSPHATE; GUANOSINE TRIPHOSPHATASE; GUANOSINE TRIPHOSPHATE; MEMBRANE RECEPTOR; MESSENGER RNA;

EID: 0031805069     PISSN: 09550674     EISSN: None     Source Type: Journal    
DOI: 10.1016/S0955-0674(98)80017-3     Document Type: Article
Times cited : (68)

References (69)
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    • The asymmetric distribution of the constituents of the Ran system is essential for transport into and out of the nucleus
    • of outstanding interest. The role of Ran-GTP distribution in nuclear transport was studied in vivo in Xenopus oocytes by collapsing the Ran-GTP gradient across the nuclear envelope. Nuclear injections of RanGAP (which activates RanGTPase), RanBP1 (which sequesters Ran-GTP) or Ran Thr24→Asn (RanT24N) (which inhibits RanGEF) functionally deplete the nucleus of Ran-GTP. These treatments block major export and import pathways, but different export pathways show distinct sensitivities to Ran-GTP depletion. Surprisingly, export of several substrates including importin α and β, transportin, HIV Rev and tRNA requires nuclear Ran-GTP but not GTP hydrolysis by Ran. This raises the possibility that the energy of GTP hydrolysis by Ran is not required for translocation through the NPC.
    • Izaurralde E, Kutay U, von Kobbe C, Mattaj IW, Görlich D. The asymmetric distribution of the constituents of the Ran system is essential for transport into and out of the nucleus. of outstanding interest EMBO J. 16:1997;6535-6547 The role of Ran-GTP distribution in nuclear transport was studied in vivo in Xenopus oocytes by collapsing the Ran-GTP gradient across the nuclear envelope. Nuclear injections of RanGAP (which activates RanGTPase), RanBP1 (which sequesters Ran-GTP) or Ran Thr24→Asn (RanT24N) (which inhibits RanGEF) functionally deplete the nucleus of Ran-GTP. These treatments block major export and import pathways, but different export pathways show distinct sensitivities to Ran-GTP depletion. Surprisingly, export of several substrates including importin α and β, transportin, HIV Rev and tRNA requires nuclear Ran-GTP but not GTP hydrolysis by Ran. This raises the possibility that the energy of GTP hydrolysis by Ran is not required for translocation through the NPC.
    • (1997) EMBO J , vol.16 , pp. 6535-6547
    • Izaurralde, E.1    Kutay, U.2    Von Kobbe, C.3    Mattaj, I.W.4    Görlich, D.5
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    • Requirement of guanosine triphosphate-bound Ran for signal-mediated nuclear protein export
    • of outstanding interest. Microinjection of RanGAP (plus or minus RanG19V) or RanT24N into nuclei of mammalian cultured cells revealed a requirement for Ran-GTP, but not GTP hydrolysis, in nuclear export mediated by leucine-rich NESs. RanG19V is a GTPase deficient mutant of Ran that is locked in the GTP bound form, whereas RanT24N is a dominant negative inhibitor of RanGEF. Differences in Ran-GTP requirements for export of various cargos were revealed by the continued export of a green fluorescent protein - NES reporter protein even when the level of nuclear Ran-GTP was reduced by inactivation of the temperature-sensitive RCC1 (RanGEF) of tsBN2 BHK cells.
    • Richards SA, Carey KL, Macara IG. Requirement of guanosine triphosphate-bound Ran for signal-mediated nuclear protein export. of outstanding interest Science. 276:1997;1842-1844 Microinjection of RanGAP (plus or minus RanG19V) or RanT24N into nuclei of mammalian cultured cells revealed a requirement for Ran-GTP, but not GTP hydrolysis, in nuclear export mediated by leucine-rich NESs. RanG19V is a GTPase deficient mutant of Ran that is locked in the GTP bound form, whereas RanT24N is a dominant negative inhibitor of RanGEF. Differences in Ran-GTP requirements for export of various cargos were revealed by the continued export of a green fluorescent protein - NES reporter protein even when the level of nuclear Ran-GTP was reduced by inactivation of the temperature-sensitive RCC1 (RanGEF) of tsBN2 BHK cells.
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    • Richards, S.A.1    Carey, K.L.2    MacAra, I.G.3
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    • CRM1 is an export receptor for leucine-rich nuclear export signals
    • of special interest. of outstanding interest. The human CRM1 protein (see Formerod 1997 [26]) is shown to be the target for leptomycin B (Wolff 1997 [23]), a cytotoxin that blocks Rev-dependent RNA export as well as export of snRNAs. Overexpression of CRM1 in oocytes reverses leptomycin-B-induced inhibition of Rev export and stimulates Rev and U snRNA export. In vitro, CRM1 binds co-operatively to both Ran-GTP and leucine-rich Rev (or protein kinase A inhibitor) NES peptides. Formation of this ternary complex is prevented in the presence of leptomycin B, supporting the conclusion that CRM1 or exportin 1 is an export receptor for leucine rich NESs.
    • of special interest Fornerod M, Ohno M, Yoshida M, Mattaj IW. CRM1 is an export receptor for leucine-rich nuclear export signals. of outstanding interest Cell. 90:1997;1051-1060 The human CRM1 protein (see Formerod 1997 [26]) is shown to be the target for leptomycin B (Wolff 1997 [23]), a cytotoxin that blocks Rev-dependent RNA export as well as export of snRNAs. Overexpression of CRM1 in oocytes reverses leptomycin-B-induced inhibition of Rev export and stimulates Rev and U snRNA export. In vitro, CRM1 binds co-operatively to both Ran-GTP and leucine-rich Rev (or protein kinase A inhibitor) NES peptides. Formation of this ternary complex is prevented in the presence of leptomycin B, supporting the conclusion that CRM1 or exportin 1 is an export receptor for leucine rich NESs.
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    • Fornerod, M.1    Ohno, M.2    Yoshida, M.3    Mattaj, I.W.4
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    • Export of importin α from the nucleus is mediated by a specific nuclear transport factor
    • of special interest. of outstanding interest. This report identifies the export receptor for importin α as cellular apoptosis susceptibility (CAS) (see also Görlich 1997 [20]), a protein previously implicated in apoptosis and cell proliferation. Using in vitro transport (digitonin-permeabilized cells) and binding assays, CAS was shown to interact with NPCs, to cross the nuclear envelope rapidly and to bind Ran-GTP and free importin α simultaneously in a highly co-operative manner. Disassembly of the trimeric importin α/CAS/Ran-GTP complex requires RanGAP and RanBP1. On the basis of these properties the authors propose that CAS is the prototype Ran-GTP-regulated shuttling export receptor, or exportin.
    • of special interest Kutay U, Bischoff FR, Kostka S, Kraft R, Görlich D. Export of importin α from the nucleus is mediated by a specific nuclear transport factor. of outstanding interest Cell. 90:1997;1061-1071 This report identifies the export receptor for importin α as cellular apoptosis susceptibility (CAS) (see also Görlich 1997 [20]), a protein previously implicated in apoptosis and cell proliferation. Using in vitro transport (digitonin-permeabilized cells) and binding assays, CAS was shown to interact with NPCs, to cross the nuclear envelope rapidly and to bind Ran-GTP and free importin α simultaneously in a highly co-operative manner. Disassembly of the trimeric importin α/CAS/Ran-GTP complex requires RanGAP and RanBP1. On the basis of these properties the authors propose that CAS is the prototype Ran-GTP-regulated shuttling export receptor, or exportin.
    • (1997) Cell , vol.90 , pp. 1061-1071
    • Kutay, U.1    Bischoff, F.R.2    Kostka, S.3    Kraft, R.4    Görlich, D.5
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    • Identification of a tRNA-specific nuclear export receptor
    • of outstanding interest. See annotation for Arts 1998 [19].
    • Kutay U, Lipowsky G, Izaurralde E, Bischoff FR, Schwarzmeier P, Hartmann E, Görlich D. Identification of a tRNA-specific nuclear export receptor. of outstanding interest Mol Cell. 1:1998;1-20 See annotation for Arts 1998 [19].
    • (1998) Mol Cell , vol.1 , pp. 1-20
    • Kutay, U.1    Lipowsky, G.2    Izaurralde, E.3    Bischoff, F.R.4    Schwarzmeier, P.5    Hartmann, E.6    Görlich, D.7
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    • Identification of a nuclear export receptor for tRNA
    • of outstanding interest. In this report and Kutay 1998 [18], exportin t was identified as a human member of the importin β family, distantly related to the yeast Los1p involved in nuclear tRNA maturation Hopper 1980 [36], Simos1996 [37]. Recombinant exportin t behaved as a nucleo - cytoplasmicshuttling protein and, unexpectedly, was found to interact directly with tRNA in a Ran-GTP-dependent manner. Injection of exportin t into nuclei of Xenopus oocytes led to specific stimulation of nuclear export of tRNAs and to the relief of the retardation of tRNA export observed in the presence of saturating amounts of tRNA.
    • Arts G-J, Fornerod M, Mattaj IW. Identification of a nuclear export receptor for tRNA. of outstanding interest Curr Biol. 8:1998;305-314 In this report and Kutay 1998 [18], exportin t was identified as a human member of the importin β family, distantly related to the yeast Los1p involved in nuclear tRNA maturation Hopper 1980 [36], Simos1996 [37]. Recombinant exportin t behaved as a nucleo - cytoplasmicshuttling protein and, unexpectedly, was found to interact directly with tRNA in a Ran-GTP-dependent manner. Injection of exportin t into nuclei of Xenopus oocytes led to specific stimulation of nuclear export of tRNAs and to the relief of the retardation of tRNA export observed in the presence of saturating amounts of tRNA.
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    • Arts G-J1    Fornerod, M.2    Mattaj, I.W.3
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    • A novel class of RanGTP binding proteins
    • of special interest. The authors describe a novel superfamily of Ran-GTP-binding proteins that includes about a dozen factors in yeast and probably even more in higher eukaryotes. The members of this superfamily (see Fornerod 1997 [26]) share with importin β an amino-terminal sequence motif that appears to account for Ran-GTP binding.
    • Görlich D, Dabrowski M, Bischoff FR, Kutay U, Bork P, Hartmann E, Prehn S, Izaurralde E. A novel class of RanGTP binding proteins. of special interest J Cell Biol. 138:1997;65-80 The authors describe a novel superfamily of Ran-GTP-binding proteins that includes about a dozen factors in yeast and probably even more in higher eukaryotes. The members of this superfamily (see Fornerod 1997 [26]) share with importin β an amino-terminal sequence motif that appears to account for Ran-GTP binding.
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    • Görlich, D.1    Dabrowski, M.2    Bischoff, F.R.3    Kutay, U.4    Bork, P.5    Hartmann, E.6    Prehn, S.7    Izaurralde, E.8
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    • Exportin 1 (Crm 1p) is an essential nuclear export factor
    • + RNA, thus indicating that the yeast mRNA export pathway may be dependent on NES-containing export adaptors (see Pasquinelli 1997 [40]).
    • + RNA, thus indicating that the yeast mRNA export pathway may be dependent on NES-containing export adaptors (see Pasquinelli 1997 [40]).
    • (1997) Cell , vol.90 , pp. 1041-1050
    • Stade, K.1    Ford, C.S.2    Guthrie, C.3    Weiss, K.4
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    • Nuclear export receptors: From importin to exportin
    • Ullmann KS, Powers MA, Forbes DJ. Nuclear export receptors: from importin to exportin. Cell. 90:1997;967-970.
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    • Ullmann, K.S.1    Powers, M.A.2    Forbes, D.J.3
  • 23
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    • Leptomycin B is an inhibitor of nuclear export: Inhibition of nucleo-cytoplasmic translocation of the human immunodeficiency virus type 1 (HIV-1) Rev protein and Rev-dependent mRNA
    • of special interest. This demonstration that leptomycin B blocks export of Rev played an important role in identification of the receptor for leucine-rich NESs.
    • Wolff B, Sanglier J-J, Wang Y. Leptomycin B is an inhibitor of nuclear export: inhibition of nucleo-cytoplasmic translocation of the human immunodeficiency virus type 1 (HIV-1) Rev protein and Rev-dependent mRNA. of special interest Chem Biol. 4:1997;139-147 This demonstration that leptomycin B blocks export of Rev played an important role in identification of the receptor for leucine-rich NESs.
    • (1997) Chem Biol , vol.4 , pp. 139-147
    • Wolff, B.1    Sanglier J-J2    Wang, Y.3
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    • The importin-beta family member Crm1p bridges the interaction between Rev and the nuclear pore complex during nuclear export
    • Neville M, Stutz F, Lee L, Davis LI, Rosbash M. The importin-beta family member Crm1p bridges the interaction between Rev and the nuclear pore complex during nuclear export. Curr Biol. 7:1997;767-775.
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    • Neville, M.1    Stutz, F.2    Lee, L.3    Davis, L.I.4    Rosbash, M.5
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    • Evidence for a role of CRM1 in signal-mediated nuclear protein export
    • Ossareh-Nazari B, Bachelerie F, Dargemont C. Evidence for a role of CRM1 in signal-mediated nuclear protein export. Science. 278:1997;141-144.
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    • Ossareh-Nazari, B.1    Bachelerie, F.2    Dargemont, C.3
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    • The human homologue of yeast CRM1 is in a dynamic subcomplex with CAN/Nup214 and a novel nuclear pore component Nup88
    • of special interest. The human CRM1 protein was found to be a nuclear and NPC-associated protein that interacts strongly with the FG-repeat region of Nup214 (see Doye1996 [5]) as well as other repeat-containing nuclear pore proteins. hCRM1 has a domain with significant homology to importin β and a group of largely uncharacterizedyeast and vertebrate proteins of similar size (110-120 kDa); this CRIME domain of several of these proteins has subsequently been shown to be involved in Ran-GTP binding (Görlich 1997 [20].
    • Fornerod M, van Deursen J, van Baal S, Reynolds A, Davis D, Murti KG, Fransen J, Grosveld G. The human homologue of yeast CRM1 is in a dynamic subcomplex with CAN/Nup214 and a novel nuclear pore component Nup88. of special interest EMBO J. 16:1997;807-816 The human CRM1 protein was found to be a nuclear and NPC-associated protein that interacts strongly with the FG-repeat region of Nup214 (see Doye1996 [5]) as well as other repeat-containing nuclear pore proteins. hCRM1 has a domain with significant homology to importin β and a group of largely uncharacterizedyeast and vertebrate proteins of similar size (110-120 kDa); this CRIME domain of several of these proteins has subsequently been shown to be involved in Ran-GTP binding (Görlich 1997 [20].
    • (1997) EMBO J , vol.16 , pp. 807-816
    • Fornerod, M.1    Van Deursen, J.2    Van Baal, S.3    Reynolds, A.4    Davis, D.5    Murti, K.G.6    Fransen, J.7    Grosveld, G.8
  • 28
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    • The K nuclear shuttling domain: A novel signal for nuclear import and nuclear export in the hnRNP K protein
    • Michael WM, Eder PS, Dreyfuss G. The K nuclear shuttling domain: a novel signal for nuclear import and nuclear export in the hnRNP K protein. EMBO J. 16:1997;3587-3598.
    • (1997) EMBO J , vol.16 , pp. 3587-3598
    • Michael, W.M.1    Eder, P.S.2    Dreyfuss, G.3
  • 29
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    • Arginine methylation facilitates the nuclear export of hnRNP proteins
    • of special interest. The predominant arginine methyltransferase of S. cerevisiae, Hmt1p, methylates both Npl3p and Hrp1p (Kessler 1997 [31]), which are shuttling hnRNP proteins involved in mRNA processing and export. In the absence of Htm1p methylation, the ability of Npl3p and Hrp1p to exit the nucleus is impaired, thus providing the first in vivo evidence of a function for hnRNP arginine methylation.
    • Shen EC, Henry MF, Weiss VH, Valentini SR, Silver PA, Lee M. Arginine methylation facilitates the nuclear export of hnRNP proteins. of special interest Genes Dev. 12:1998;679-691 The predominant arginine methyltransferase of S. cerevisiae, Hmt1p, methylates both Npl3p and Hrp1p (Kessler 1997 [31]), which are shuttling hnRNP proteins involved in mRNA processing and export. In the absence of Htm1p methylation, the ability of Npl3p and Hrp1p to exit the nucleus is impaired, thus providing the first in vivo evidence of a function for hnRNP arginine methylation.
    • (1998) Genes Dev , vol.12 , pp. 679-691
    • Shen, E.C.1    Henry, M.F.2    Weiss, V.H.3    Valentini, S.R.4    Silver, P.A.5    Lee, M.6
  • 30
    • 0031929940 scopus 로고    scopus 로고
    • A specific subset of SR proteins shuttles continuously between the nucleus and the cytoplasm
    • of special interest. This study shows that different members of the family of mammalian Ser-Arg (SR) repeat proteins, which are nuclear phosphoproteins required for constitutive pre-mRNA splicing, have unique intracellular transport properties. Shuttling of SF2/ASF depends both the carboxy-terminal arginine/serine-rich (RS) domains and stable RNA binding, as revealed by the requirement for a functional RNA recognition motif, RRM1; moreover, hyperphosphorylation of the RS domain leads to increased cytoplasmic accumulation of SF2/ASF. Thus, SR shuttling proteins, such as SF2/ASF and SRp20, may have roles not only in pre-mRNA splicing, but also in mRNA transport and related cytoplasmic events.
    • Caceres JF, Screaton GR, Krainer AR. A specific subset of SR proteins shuttles continuously between the nucleus and the cytoplasm. of special interest Genes Dev. 12:1998;55-66 This study shows that different members of the family of mammalian Ser-Arg (SR) repeat proteins, which are nuclear phosphoproteins required for constitutive pre-mRNA splicing, have unique intracellular transport properties. Shuttling of SF2/ASF depends both the carboxy-terminal arginine/serine-rich (RS) domains and stable RNA binding, as revealed by the requirement for a functional RNA recognition motif, RRM1; moreover, hyperphosphorylation of the RS domain leads to increased cytoplasmic accumulation of SF2/ASF. Thus, SR shuttling proteins, such as SF2/ASF and SRp20, may have roles not only in pre-mRNA splicing, but also in mRNA transport and related cytoplasmic events.
    • (1998) Genes Dev , vol.12 , pp. 55-66
    • Caceres, J.F.1    Screaton, G.R.2    Krainer, A.R.3
  • 31
    • 0030803670 scopus 로고    scopus 로고
    • Hrp1, a sequence-specific RNA binding protein that shuttles between the nucleus and the cytoplasm, is required for mRNA 3′-end formation in yeast
    • of special interest. This study identifies Hrp1p as an essential component of the yeast 3′ polyadenylation machinery and demonstrates that this specific RNA binding protein, like Npl3p, shuttles between the nucleus and the cytoplasm. Thus, Hrp1p provides a direct link between 3′ end processing and nuclear export of mRNA. The HRP1 gene was originally isolated as a suppressor of a temperature-sensitive allele of NPL3, a gene encoding a protein involved in mRNA export (Shen 1998 [29]).
    • Kessler MM, Henry MF, Shen E, Zhao J, Gross S, Silver PA, Moore CL. Hrp1, a sequence-specific RNA binding protein that shuttles between the nucleus and the cytoplasm, is required for mRNA 3′-end formation in yeast. of special interest Genes Dev. 11:1997;2545-2556 This study identifies Hrp1p as an essential component of the yeast 3′ polyadenylation machinery and demonstrates that this specific RNA binding protein, like Npl3p, shuttles between the nucleus and the cytoplasm. Thus, Hrp1p provides a direct link between 3′ end processing and nuclear export of mRNA. The HRP1 gene was originally isolated as a suppressor of a temperature-sensitive allele of NPL3, a gene encoding a protein involved in mRNA export (Shen 1998 [29]).
    • (1997) Genes Dev , vol.11 , pp. 2545-2556
    • Kessler, M.M.1    Henry, M.F.2    Shen, E.3    Zhao, J.4    Gross, S.5    Silver, P.A.6    Moore, C.L.7
  • 32
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    • An RNA-export mediator with an essential nuclear export signal
    • of special interest. This and related papers [33-35] describe soluble adaptors that participate in mRNA export.
    • Murphy R, Wente S. An RNA-export mediator with an essential nuclear export signal. of special interest Nature. 383:1996;36-357 This and related papers [33-35] describe soluble adaptors that participate in mRNA export.
    • (1996) Nature , vol.383 , pp. 36-357
    • Murphy, R.1    Wente, S.2
  • 33
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    • Yeast heat shock mRNAs are exported through a distinct pathway defined by Rip1p
    • of special interest. This and related papers [32-35] describe soluble adaptors that participate in mRNA export.
    • Saavedra CA, Hammell CM, Heath CV, Cole CN. Yeast heat shock mRNAs are exported through a distinct pathway defined by Rip1p. of special interest Genes Dev. 11:1997;2845-2856 This and related papers [32-35] describe soluble adaptors that participate in mRNA export.
    • (1997) Genes Dev , vol.11 , pp. 2845-2856
    • Saavedra, C.A.1    Hammell, C.M.2    Heath, C.V.3    Cole, C.N.4
  • 34
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    • The yeast nucleoporin Rip1p contributes to multiple export pathways with no essential role for its FG-repeat region
    • of special interest. This and related papers [32-35] describe soluble adaptors that participate in mRNA export.
    • Stutz F, Kantor J, Zhang D, McCarthy T, Neville M, Rosbash M. The yeast nucleoporin Rip1p contributes to multiple export pathways with no essential role for its FG-repeat region. of special interest Genes Dev. 11:1997;2857-2868 This and related papers [32-35] describe soluble adaptors that participate in mRNA export.
    • (1997) Genes Dev , vol.11 , pp. 2857-2868
    • Stutz, F.1    Kantor, J.2    Zhang, D.3    McCarthy, T.4    Neville, M.5    Rosbash, M.6
  • 35
    • 0030911425 scopus 로고    scopus 로고
    • Mex67p, a novel factor for nuclear mRNA export binds to both poly(A)+ RNA and nuclear pores
    • of special interest. This and related papers [32-35] describe soluble adaptors that participate in mRNA export.
    • Segref A, Sharma K, Doye V, Hellwig A, Huber J, Lührmann R, Hurt E. Mex67p, a novel factor for nuclear mRNA export binds to both poly(A)+ RNA and nuclear pores. of special interest EMBO J. 16:1997;3256-3271 This and related papers [32-35] describe soluble adaptors that participate in mRNA export.
    • (1997) EMBO J , vol.16 , pp. 3256-3271
    • Segref, A.1    Sharma, K.2    Doye, V.3    Hellwig, A.4    Huber, J.5    Lührmann, R.6    Hurt, E.7
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    • Processing of intervening sequences: A new yeast mutant fails to excise intervening sequences from precursor tRNAs
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    • The constitutive transport element (CTE) from simian retrovirus-1, unlike the HIV-1 RRE-Rev system, utilizes factors required for the export of cellular mRNAs
    • of special interest. See annotation to Pasquinelli 1997 [39].
    • Saavedra C, Felber B, Izaurralde E. The constitutive transport element (CTE) from simian retrovirus-1, unlike the HIV-1 RRE-Rev system, utilizes factors required for the export of cellular mRNAs. of special interest Curr Biol. 7:1997;619-628 See annotation to Pasquinelli 1997 [39].
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    • The constitutive transport element (CTE) of Mason-Pfizer monkey virus (MPMV) accesses a cellular mRNA export pathway
    • of special interest. Using Xenopus oocytes, the constitutive transport element (CTE) and Rev - RRE export systems, both of which function in export of intron-containing viral mRNAs, were shown to require different saturable export factors. CTE RNA very effectively titrates certain factors essential for mRNA export; however, reciprocal competition experiments with high amounts of hnRNP A1 protein or DHFR mRNA do not block CTE RNA export, indicating that additional factors can become limiting when mRNA export is saturated. The CTE has a pseudo-twofold symmetric structure, either half of which is sufficient to support export of an intron-lariat and to titrate mRNA export factors.
    • Pasquinelli AE, Ernst RK, Lund E, Grimm D, Zapp ML, Rekosh D, Hammarskjöld M-L, Dahlberg JE. The constitutive transport element (CTE) of Mason-Pfizer monkey virus (MPMV) accesses a cellular mRNA export pathway. of special interest EMBO J. 16:1997;7500-7510 Using Xenopus oocytes, the constitutive transport element (CTE) and Rev - RRE export systems, both of which function in export of intron-containing viral mRNAs, were shown to require different saturable export factors. CTE RNA very effectively titrates certain factors essential for mRNA export; however, reciprocal competition experiments with high amounts of hnRNP A1 protein or DHFR mRNA do not block CTE RNA export, indicating that additional factors can become limiting when mRNA export is saturated. The CTE has a pseudo-twofold symmetric structure, either half of which is sufficient to support export of an intron-lariat and to titrate mRNA export factors.
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    • Pasquinelli, A.E.1    Ernst, R.K.2    Lund, E.3    Grimm, D.4    Zapp, M.L.5    Rekosh, D.6    Hammarskjöld M-L7    Dahlberg, J.E.8
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    • of outstanding interest Pasquinelli AE, Powers MA, Lund E, Forbes D, Dahlberg JE. Inhibition of mRNA export in vertebrate cells by nuclear export signal conjugates. of special interest Proc Natl Acad Sci USA. 94:1997;14394-14399 Rev and protein kinase A inhibitor (PKI) NES peptide conjugates were shown to be effective inhibitors of both U snRNA and mRNA export in micro-injected Xenopus oocytes. This is in agreement with the requirement for yeast Xpo1p in mRNA export (Stade 1997 [21]), but it is in contrast to other studies linking only the Rev and U snRNA and 5S rRNA export pathways (Fischer 1995 [41]). Control experiments indicate that the differences in inhibitory activities of various preparations of Rev NES-conjugates most probably reflect differences the methods of peptide conjugation.
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    • RanBP1 is crucial for the release of RanGTP from importin β-related nuclear transport factors
    • of special interest. The authors describe three cases in which RanBP1 stimulated, by more than two orders of magnitude, the removal of Ran-GTP from transport receptor/Ran-GTP/RanBP1 complexes, leading to their disassembly, and to the hydrolysis of GTP in the presence of RanGAP. The efficient release of importin β from Ran-GTP was found to require importin α in addition to RanBP1.
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    • Ranunassisted nuclear migration of a 97-kD component of the nuclear pore-targeting complex
    • of outstanding interest. This study examined the transport behavior of PTAC97 (importin β) in living cells and digitonin-permeabilized cells when it was not carrying the importin α/NLS-cargo complex. Surprisingly, importin β was rapidly translocated into the nucleus in a saturable, but RanGTPase independent manner. The use of deletion mutants, which lacked the ability to bind importin α, Ran-GTP or both, revealed that the NPC binding domain of importin β was both necessary and sufficient for translocation through the NPC. Moreover, a dominant-negative variant of Ran, defective in GTP hydrolysis, did not inhibit import of importin β in living cells (see also Nakielny 1997 [51].
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    • Import and export of the nuclear protein import receptor transportin by a mechanism independent of GTP hydrolysis
    • of outstanding interest. Using a similar approach as the study in Kose 1997 [50], import and export of transportin (the import receptor for hnRNP A1) were shown to be inhibited by low temperature, but not by the non-hydrolyzable GTP analogue GMP - PNP. These results indicate that temperature-dependent transport through the NPC can be uncoupled from the RanGTPase cycle and can occur without GTP hydrolysis.
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