Nonrepetitive DNA Sequence Representation in Sea Urchin Embryo Messenger RNA

Goldberg, Robert B.; Galau, Glenn A.; Britten, Roy J.; Davidson, Eric H.

doi:10.1073/pnas.70.12.3516

Cited by 80 publications

(34 citation statements)

References 17 publications

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“…The results also suggest strongly that in each case mRNA is transcribed preferentially from the single-copy fraction of the DNA. This is in agreement with previous work from this and from other laboratories on mRNA sequences in various organisms (20)(21)(22)(23). This result is particularly striking in Triturus, since the majority of the genome consists of repeated DNA.…”

supporting

confidence: 82%

“…3 and 6 strongly suggest that the basic transcription patterns in Xenopus and Triturus ovaries are very similar. In both cases, mRNA is transcribed preferentially from the single-copy fraction of the genome, in agreement with previous results on other systerns (20)(21)(22)(23)(24). Second, the number of different RNA species present in the ovaries of the two organisms is similar.…”

supporting

confidence: 81%

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Analysis of the C-Value Paradox by Molecular Hybridization

Rosbash

Ford

Bishop

1974

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

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Poly(A)-containing RNA was isolated from ovaries of Xenopus laevis laevis and Triturus cristatus carnifex and used as a template for the synthesis of radioactive complementary DNA with RNA-dependent DNA polymerase. When annealed with an excess of homologous DNA, the complementary DNA is rendered double-stranded with kinetics that suggest that the coding sequences are singlecopy in both these organisms. In Triturus, these sequences are distinct from the majority of the genome, which consists of repeated sequences, and distinct from the ribosomal cistrons, which are present in proportion to the increase in C-value relative to the Xenopus genome. Moreover, the number of different poly(A)-containing molecules in the ovary (sequence complexity) is the same in Xenopus and in Triturus.There exists an enormous variation in C-value (DNA content per haploid genome) among eukaryotic organisms (1 (3,4).On the basis of this range of DNA values and cytological evidence from various sources, Callan proposed that chromosomal DNA, and in particular genetically significant DNA, is arranged in tandemly repeated sequences (5). These repeated sequences consist of a "master" sequence and "slave" sequences; the "master" sequence would be subject to mutational and recombinational events while the "slave" sequences would be corrected once per life-cycle to reflect the sequence of the "master" sequence. The formation of extended lampbrush loops during meiotic prophase was proposed as the stage at which this correction process takes place.In washed in Barth X medium. Nucleic acid was prepared by the method of Kirby, with the addition of 0.5% sodium lauryl sulfate to the aqueous phase, as described (7). Highmolecular-weight RNA was prepared by homogenizing the pelleted nucleic acid in 3 M NaOAc (pH 6) to solubilize DNA, transfer RNA, and 5S RNA (8). The RNA precipitate was resuspended in "binding" buffer [0.4 M NaCl, 1 mM EDTA, 0.1% sodium lauryl sulfate, 10 mM Tris * HCl (pH 7.4), 10% glycerol] and passed over oligo(dT)-cellulose (9). The poly-(A)-containing RNA was eluted in "elution" buffer [10 mM Tris * HCl pH 7.4, 0.1% sodium lauryl sulfate], and the RNA spectrum was measured in the Unicam SP800A spectrophotometer. The amount of poly(A) in this eluate was routinely assayed by hybridization with radioactive poly(U), as described (7). By comparing the amount of poly (A) (Fig. 1). About 75% of

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supporting

confidence: 82%

supporting

confidence: 81%

Analysis of the C-Value Paradox by Molecular Hybridization

Rosbash

Ford

Bishop

1974

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

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“…This observation, first made in HeLa cells (1), has been repeated for cells of a number of different organisms (2)(3)(4)(5)(6)(7)(8)(9)(10). The broad abundance distribution of mRNA sequences presumably results from fundamental regulatory mechanisms which adjust the rate of production of individual proteins.…”

Section: Introductionmentioning

confidence: 99%

Messenger RNA abundance and lifetime: a correlation in Drosophila cells but not in HeLa

1978

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Messenger RNA in eukaryotes is comprised of several abundance classes. Also, the decay of these unstable molecules shows at least two very different lifetimes. Two independent techniques are used here to examine the relation between message abundance and lifetime in cell lines from very different organisms. The methods give consistent results for each cell line; however, the two cell types show very different results. In Drosophila cells, slowly decaying sequences fall in the abundant class while scarce sequences turn over rapidly. In contrast, in HeLa cells the abundant and scarce message classes are each comprised of long-and short-lived molecules.

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“…RNA isolation was carried out using the protocol of Schmidt et al (24) Analysis of Leaf Polyribosomes. Polyribosomes were isolated from leaf base (0-4 cm) and tip (7 cm to tip) samples using the protocol of Goldberg et al (9) with the following modifications. The frozen tissue was initially ground to a fine powder in a dry ice-chilled coffee grinder before being suspended in extraction buffer.…”

mentioning

confidence: 99%

Photosynthetic Gene Expression and Cellular Differentiation in Developing Maize Leaves

Martineau¹,

Taylor²

1985

Plant Physiol.

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We have exploited the positional gradient of cellular differentiation in Zea mays leaves to study the accumulation of mRNAs encoding subunits ofthe two COrfixing enzymes and the major chlorophyll-binding protein. These multi-step scheme of CO2 fixation called C4 photosynthesis. PEPCase3, the enzyme responsible for initially fixing the atmospheric CO2 in maize, is found only in mesophyll cells (22). The four-carbon acid resulting from this reaction is shuttled to bundle sheath cells where it is decarboxylated, supplying CO2 to carry out the reductive pentose phosphate cycle, the initial product of which is a three-carbon compound, as in C3 plants (4). RuBPCase is found only in the chloroplasts of bundle sheath cells (12). This compartmentalization of the carboxylases confers photosynthetic advantage to C4 plants under conditions of high temperature and light intensity (11). In addition to these enzymic differences between the mesophyll and bundle sheath cells of C4 plants, there are differences in the chloroplast membrane proteins of the two cell types. Membrane proteins from maize mesophyll cell chloroplasts are essentially identical to those of several C3 plants, containing polypeptides responsible for the light-harvesting and energy-transduction reactions of photosynthesis (2). The bundle sheath cell chloroplasts, however, contain little if any of the PSII protein complex and its associated LHCP (1; Schuster G, I Ohad, B Martineau, WC Taylor, unpublished). We have outlined the patterns of mRNA accumulation for four of the compartmentalized polypeptides related to C4 photosynthesis by using hybridization with DNA probes to measure accumulation of RuBPCase (LSu and SSu), PEPCase, and LHCP mRNAs in cells of increasing age and stage of differentiation. We compare our patterns of mRNA accumulation for these proteins to the patterns of their polypeptide accumulation established by Mayfield and Taylor (17) in a similar study. We have monitored leaf morphological development by using a cytological marker to indicate fully differentiated vascular bundles, again taking advantage of the gradient of cellular development in the young leaves we have chosen to study.MATERIALS AND METHODS Plant Material. Seeds from an inbred line of Zea mays (B73, a gift of Pioneer Hi-Bred International, Johnston, IA) were planted in a 2:1 mixture of soil and sand, covered with 2 cm of vermiculite, and germinated and grown in a growth chamber. Periods of 16 h light (4 x 104 lux) and 8 h dark, at 25°C, were used. Plants were harvested 10 to 14 d after germination when the third leaf to emerge from the coleoptile was 12 to 16 cm long. A third leaf of this age has not yet developed a ligule, the structure which delineates the leaf blade from its sheath. The coleoptile and first and second leaves were removed from each plant. The third leaf was cut at points 1, 2, 4, 6, and 8 cm from the leaf base. The six resulting leaf segments, representing populations of cells of the same relative age and stage of differentiation, were separated and...

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Nonrepetitive DNA Sequence Representation in Sea Urchin Embryo Messenger RNA

Cited by 80 publications

References 17 publications

Analysis of the C-Value Paradox by Molecular Hybridization

Analysis of the C-Value Paradox by Molecular Hybridization

Messenger RNA abundance and lifetime: a correlation in Drosophila cells but not in HeLa

Photosynthetic Gene Expression and Cellular Differentiation in Developing Maize Leaves

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