Differential changes of nuclear-envelope-associated enzyme activities involved in nucleocytoplasmic mRNA transport in the developing rat brain and liver

Schröder, Heinz C.; Becker, Roger; Bachmann, Michael; Gramzow, Monika; Sève, Annie-Pierre; Monsigny, Michel; Müller, Wernér E.G.

doi:10.1016/0167-4781(86)90013-8

Cited by 16 publications

(8 citation statements)

References 49 publications

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“…However, after addition of 6 mg/ml of total dialyzed L-cell cytosol protein the ATP-promoted efflux of rapidly labeled RNA from Lcell nuclei was raised to 17.2% total nuclear radioactivity (incubation period, 30 min) and thus reached a similar extent as that from nuclear matrices. This finding is in agreement with our previous results [21]. Interestingly, the amount of RNA released by cytochalasin B from the isolated nuclei was not changed in the presence of cytosolic protein (3.7% total nuclear radioactivity as compared with 3.6%, measured in protein-free transport medium; see Fig.…”

Section: Resultssupporting

confidence: 83%

“…The influence of agents affecting actin assembly/disassembly on the release and translocation steps of nucleo- cytoplasmic RNA transport was studied using the following two in vitro systems: (a) isolated hen oviduct cell nuclei and nuclear matrices for measurement of the release of a highabundance mRNA (ovalbumin mRNA representing about 50% of the total mRNA population in the mature oviduct) [21]; (b) isolated nuclei and nuclear matrices from L-cells for measurements of the release of rapidly labeled, total RNA (primarily mRNA under the labeling conditions chosen [9, As shown in Fig. 1, all of the ovalbumin sequence containing RNA bands that were detected in whole oviduct cell nuclei with the pOV9.8 probe (lane a) were also found to be present in the nuclear matrix fraction (lane b).…”

Section: Resultsmentioning

confidence: 99%

“…Under these conditions, radioactivity is predominantly incorporated into the mRNA and its precursors [21].…”

Section: Cell Culturementioning

confidence: 99%

“…The conditions for RNA efflux measurements have been described earlier [21,25,261. Briefly, nuclei (from L-cells prelabeled with [3H]uridine or from hen oviduct) were washed twice in transport medium [50 mM Tris/HCl, pH 7.6,25 mM KCl, 250 mM sucrose, 2.5 mM MgC12, 0.3 mM MnCI2, 0.5 mM CaCI,, 5 mM 2-mercaptoethanol, 5 mM spermidine HC1, and lo3 units/ml of RNasin (used if Northern and dotblot hybridization experiments were performed) or 0.3 mg/ml of heat-treated yeast RNA (measurement of efflux of rapidly labeled RNA)] and then resuspended in transport medium to a concentration of about 2.5 x 106/ml.…”

Section: Rna Ejjlux From Isolated Nucleimentioning

confidence: 99%

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Cytochalasin B selectively releases ovalbumin mRNA precursors but not the mature ovalbumin mRNA from hen oviduct nuclear matrix

Schröder

Trölltsch

Wenger

et al. 1987

European Journal of Biochemistry

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Hen oviduct nuclear matrix-bound mature ovalbumin mRNA is released from the matrix in the presence of ATP, while the ovalbumin mRNA precursors remain bound to this structure. Detachment of the mature mRNA from the matrix by ATP as well as ATP-dependent efflux of mRNA from isolated nuclei were found to be inhibited by cytochalasin B. On the other hand, in the absence of ATP, cytochalasin B exclusively caused the release (and nucleocytoplasmic efflux) of the ovalbumin messenger precursors, but not of the mature mRNA. After cytochalasin B treatment, actin could be detected in the matrix supernatant. Phalloidin which stabilizes actin filaments did not cause RNA liberation in the absence of ATP, but inhibited the ATP-induced detachment of mature mRNA. RNA release was also achieved with a monoclonal antibody against actin but not with monoclonal antibodies against tubulin and intermediate filaments. These results suggest that actin-containing filaments are involved in the restriction of immature messengers to the cell nucleus.It is well established that the immature precursors of cytoplasmic mRNAs are restricted to the nucleus [l, 21. Therefore, transport of mRNA from nucleus to cytoplasm is assumed to be associated with a selection of exclusively mature mRNA molecules [2]. In the nucleus the mRNA precursors were found almost quantitatively attached to an internal nuclear structure that has been termed the nuclear matrix [3-91. In contrast to the immature mRNA, only about 30 -50% of the mature mRNA has been shown to be bound to this structure [8, 91. This matrix-bound fraction of the mature mRNA displayed no difference in its binding strength compared with the hnRNA [S, 91. Because the mRNA seems to be largely associated with intracellular structures, both in the nucleus and in the cytoplasm [lo-121, nucleocytoplasmic mRNA transport is considered as a solid-phase, multistep process [2, 131 comprising (a) the release of the mRNA from the matrix, (b) the translocation of the probably carrier-bound mRNA through a nuclear envelope pore complex, and (c) the binding of the transported mRNA to the cytoskeleton. The translocation step has been shown to be mediated by a nuclear envelope nucleoside triphosphatase [13, 141 which is stimulated by the 3' poly(A) sequence of mRNA [15]. This enzyme has been solubilized from rat liver envelopes by Triton X-100 [16] and purified to homogeneity [17]. Recently we demonstrated that the selection of the mature mRNAs for transport into the cytoplasm occurs during their release from the matrix [9]. The

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Section: Resultssupporting

confidence: 83%

Section: Resultsmentioning

confidence: 99%

“…Under these conditions, radioactivity is predominantly incorporated into the mRNA and its precursors [21].…”

Section: Cell Culturementioning

confidence: 99%

Section: Rna Ejjlux From Isolated Nucleimentioning

confidence: 99%

See 2 more Smart Citations

Cytochalasin B selectively releases ovalbumin mRNA precursors but not the mature ovalbumin mRNA from hen oviduct nuclear matrix

Schröder

Trölltsch

Wenger

et al. 1987

European Journal of Biochemistry

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“…Preincubation with different concentrations of Gerardia lectin was done for 10 min at 22°C in transport medium without ATP and the ATP-generating system. RNA in the postnuclear supernatant was bound to a poly(U) filter (messenger-activated paper) and determined as described [28]. In the absence of ATP in the transport medium, the mRNA efflux rate was 0.3 x lo4 dpm (lo6 nuclei)-' (30 min)-'.…”

Section: Fractionation Of Nucleimentioning

confidence: 99%

A d-mannose-specific lectin from Gerardia savaglia that inhibits nucleocytoplasmic transport of mRNA

Kljajić¹,

Schröder

Rottmann

et al. 1987

Eur J Biochem

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A new lectin has been isolated from the coral Gerardia savaglia by affinity chromatography, using locust gum as an absorbent, and d‐mannose as eluant. Final purification was achieved by Bio‐Gel P300 gel filtration. The agglutinin is a protein composed of two polypeptide chains with a Mr of 14800; the two subunits are not linked by disulfide bond(s). The isoelectric point is 4.8, the amino acid composition is rich in the acidic amino acids aspartic acid and glutamic acid. The absorption maximum for the protein was at 276 nm; with a molar absorption coefficient of 1.27 × 105 M−1 cm−1. The lectin precipitated erythrocytes from humans (A, B and O), sheep, rabbit and carp with a titer between 25 and 1010; the affinity constant for lectin binding to sheep red blood cells was 2.8 × 108 M−1 and the number of binding sites, 3.2 × 105/cell. Ca2+ ions are required for full activity; the pH optimum lies in the range between 6 and 11. Inhibition experiments revealed that the lectin is specific for d‐mannose. The lectin is mitogenic only for those spleen lymphocytes from mice which had been activated by lipopolysaccharide. An interesting feature of this lectin is its ability to bind to glycoproteins present in nuclei from CV‐1 monkey kidney cells. The fluorescein‐isothiocyanate‐labelled lectin reacted with six polypeptides in the nuclear envelope from rat liver (Mr 190000, 115000, 80000, 62000, 56000 and 42000) and with two polypeptides in the nuclear matrix or pore complex lamina fraction (Mr 190000 and 62000). The lectin inhibited the nuclear envelope mRNA translocation system in vitro. It is suggested that this effect is due to an interaction of the lectin with the nuclear glycoproteins gp190 and/or gp62.

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Regulation and Pathologies of Nucleocytoplasmic Transport

Agutter

Taylor²

1996

The Meaning of Nucleocytoplasmic Transport

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Differential changes of nuclear-envelope-associated enzyme activities involved in nucleocytoplasmic mRNA transport in the developing rat brain and liver

Cited by 16 publications

References 49 publications

Cytochalasin B selectively releases ovalbumin mRNA precursors but not the mature ovalbumin mRNA from hen oviduct nuclear matrix

Cytochalasin B selectively releases ovalbumin mRNA precursors but not the mature ovalbumin mRNA from hen oviduct nuclear matrix

A d-mannose-specific lectin from Gerardia savaglia that inhibits nucleocytoplasmic transport of mRNA

Regulation and Pathologies of Nucleocytoplasmic Transport

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