Developmental changes in the composition of polyadenylated RNA isolated from free and membrane-bound polyribosomes of the rat forebrain, analysed by translation <i>in vitro</i>

Hall, Christine; Lim, Louis

doi:10.1042/bj1960327

Cited by 25 publications

(27 citation statements)

References 48 publications

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“…The size of synthesized proteins varies with the corresponding increase in size of RNA as estimated by sedimentation. The size distributions of translation products of both this size-fractionated RNA and total unfractionated RNA (results not shown) were very similar to that for the corresponding fractions of rat brain RNA (Hall & Lim, 1981).…”

Section: Protein Synthesissupporting

confidence: 53%

“…Protease digestion was performed in situ during re-electrophoresis of the excised proteins as described by Cleveland et al (1977). (Hall & Lim, 1981 (Hall & Lim, 1981) and under denaturing conditions with 85% (v/v) formamide (Elliott et al, 1980). The composition of translation products from the serial gradient fractions is also shown in Fig.…”

Section: Protein Synthesismentioning

confidence: 99%

“…After brief warming to 65°C, RNA was further purified by phenol extraction. Ethanol-precipitated RNA was dissolved, applied to an oligo(dT)-cellulose column and chromatographed as described previously (Hall & Lim, 1981). Unbound RNA was washed from the column by high-salt buffer [0.5M-NaCl/1 mM-EDTA/l0mM-Tris/HCl (pH 7.5)/0.1% SDS].…”

mentioning

confidence: 99%

“…Serial fractions (0.2 ml) were collected and precipitated with ethanol as described previously (Hall & Lim, 1981).…”

mentioning

confidence: 99%

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Alterations in the relative amounts of specific mRNA species in the developing human brain in Down's syndrome

Whatley¹,

Hall²,

Davison³

et al. 1984

Biochemical Journal

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Total cellular polyadenylated RNA [poly(A)+ RNA] was prepared after guanidinium thiocyanate extraction of frozen brain tissue from age-matched normal and Down'ssyndrome (trisomy 21) human foetuses. Poly(A)+ RNA populations were analysed by translation in vitro, followed by two-dimensional gel analysis by using both isoelectric focusing (ISODALT system) and non-equilibrium pH-gradient electrophoresis (BASODALT system) as the first-dimension separation. The relative concentrations of poly(A)+ RNA species coding for seven translation products were significantly altered in Down's syndrome, as determined by both visual comparisons of translation-product fluorograms from normal and Down's-syndrome samples and by quantitative radioactivity determination of individual translation products. The relative concentrations of mRNA species coding for two proteins (68 kDa and 49 kDa) were increased in Down's syndrome and may represent genes located on chromosome 21. The relative concentrations of mRNA species coding for five proteins (37 kDa, 35 kDa, 25.5 kDa, 24.5 kDa, 23 kDa) were decreased in Down's syndrome, these probably representing secondary effects of the trisomy. Six Down'ssyndrome-linked translation products (49 kDa, 37 kDa, 33 kDa, 25.5 kDa, 24.5 kDa, 23 kDa) did not migrate with appreciable amounts of cellular proteins on two-dimensional gels and hence may represent either proteins of high turnover rates or those that are post-translationally modified in vivo. One translation product (68 kDa) comigrated with a major cellular protein species, which was identified as a 68 kDa microtubule-associated protein by limited peptide mapping. The significance of these changes is discussed in relation to the mechanisms whereby the Down's-syndrome phenotype is expressed in the human brain.

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Section: Protein Synthesissupporting

confidence: 53%

Section: Protein Synthesismentioning

confidence: 99%

mentioning

confidence: 99%

“…Serial fractions (0.2 ml) were collected and precipitated with ethanol as described previously (Hall & Lim, 1981).…”

mentioning

confidence: 99%

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Alterations in the relative amounts of specific mRNA species in the developing human brain in Down's syndrome

Whatley¹,

Hall²,

Davison³

et al. 1984

Biochemical Journal

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Translation in vitro of membrane-bound polyribosomal mRNAs from rat brain has shown several to be developmentally regulated [Hall & Lim (1981) Biochem. J.

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confidence: 99%

Expression and developmental regulation of two unique mRNAs specific to brain membrane-bound polyribosomes

Hall¹,

Lowndes²,

Leung³

et al. 1987

Biochemical Journal

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Translation in vitro of membrane-bound polyribosomal mRNAs from rat brain has shown several to be developmentally regulated [Hall & Lim (1981) Biochem. J. 196,[327][328][329][330][331][332][333][334][335][336]. Here we describe the isolation and characterization of cDNAs corresponding to two such brain mRNAs. One cDNA (M444) hybridselected a 0.95 kb mRNA directing the synthesis in vitro of a 21 kDa pl-6.3 polypeptide, which was processed in vitro by microsomal membranes. A second cDNA (M1622) hybridized to a 2.2 kb mRNA directing the synthesis of a 55 kDa pl-5.8 polypeptide. Both mRNAs were specific to membrane-bound polyribosomes. Restriction maps of the corresponding genomic DNA sequences are consistent with both being single copy. The two mRNAs were present in astrocytic and neuronal cultures, but not in liver or spleen or in neuroblastoma or glioma cells. The two mRNAs were differently regulated during brain development. In the developing forebrain there was a gradual and sustained increase in M444 mRNA during the first 3 weeks post partum, whereas M 1622 mRNA appeared earlier and showed no further increase after day 10. In the cerebellum the developmental increase in M444 mRNA was biphasic. After a small initial increase there was a decrease in this mRNA at day 10, coincident with high amounts of M1622 mRNA. This was followed by a second, larger, increase in M444 mRNA, when amounts of M1622 mRNA were constant. The contrasting changes in these two mRNAs in the developing cerebellum are of particular interest, since they occur during an intensive period of cell proliferation, migration and altering neural connectivity. As these mRNAs are specific to differentiated neural tissue, they represent useful molecular markers for studying brain differentiation.

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The Effect of Trisomy-21 (Down’s Syndrome) on Brain Transcription

Lim¹,

Hall²,

Leung³

et al. 1986

Topics in the Neurosciences

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Developmental changes in the composition of polyadenylated RNA isolated from free and membrane-bound polyribosomes of the rat forebrain, analysed by translation in vitro

Cited by 25 publications

References 48 publications

Alterations in the relative amounts of specific mRNA species in the developing human brain in Down's syndrome

Alterations in the relative amounts of specific mRNA species in the developing human brain in Down's syndrome

Expression and developmental regulation of two unique mRNAs specific to brain membrane-bound polyribosomes

The Effect of Trisomy-21 (Down’s Syndrome) on Brain Transcription

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