Messenger RNA sequence complexity and homology in developmental stages of Drosophila

Lévy, Laurence; Manning, Jerry E.

doi:10.1016/0012-1606(81)90243-8

Cited by 40 publications

(19 citation statements)

References 22 publications

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“…It is evident that, in other organisms, RNA transcripts can be found in the nucleus for genes that are not expressed in that particular developmental stage or tissue but are expressed in other stages or tissue types (33,34). Our previous results (17), which indicate little qualitative change in the polysomal mRNA population in adults, larvae, and pupae, do not support this interpretation. However, it should be noted that these stages represent a limited sampling ofthe development ofDrosophila.…”

Section: Resultsmentioning

confidence: 65%

“…Labeled single-copy DNA prepared in this manner had an average single-strand length of 210 nucleotides and a specific activity of 6-7 X 10' cpm/,ug. Approximately 90% of this DNA reassociated when mixed in trace quantities with excess amounts of total D. melanogaster nuclear DNA. mDNA from third-instar larvae was prepared as described (17).…”

Section: Methodsmentioning

confidence: 99%

“…To achieve this, poly(A)+mRNA (10 ,ug) was hybridized in >100-fold sequence excess with [3H]mDNA (5 X 10-3 ,g). DNA in hybrid form was separated from unhybridized DNA by centrifugation to equilibrium twice in a neutral CsCl gradient as described (17). A band of single-stranded, noncomplementary DNA was isolated from the gradient, and contaminating RNA was removed by treatment with 0.1 M NaOH at 60°C for 30 …”

Section: Methodsmentioning

confidence: 99%

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Nonadenylylated mRNA is present as polyadenylylated RNA in nuclei of Drosophila.

Zimmerman¹,

Fouts²,

Lévy³

et al. 1982

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

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The sequence complexity of nuclear total RNA and nuclear poly(A)+RNA from Drosophilathird-instar larvae was determined by hybridization of these RNAs to labeled single-copy DNA. At saturation, the nuclear poly(A)+-and total RNA hybridized to 11% and 22.5% of the single-copy DNA, respectively. The increase in complexity ofnuclear total RNA over that observed for nuclear poly(A)+RNA indicates the presence of a discrete class of nonadenylylated nuclear RNA molecules. The relationship between DNA sequences coding for nuclear RNA and mRNA was then determined by hybridization of nuclear total and poly(A)+RNA to DNA enriched for mRNA coding sequences. The results ofthese studies show that those single-copy DNA sequences that are represented in either the poly(A)+-or poly(A)-mRNA population are transcribed into RNA molecules that appear in the nuclear poly(A)+RNA population.It has been proposed that high molecular weight nuclear RNA serves several functions within the eukaryotic cell, the most widely accepted ofwhich is as a precursor to cytoplasmic mRNA (1). Although little direct evidence for this role existed for several years, recent advances in molecular techniques have resulted in the identification and characterization ofnuclear precursors for several specific mRNAs (2-5). In these studies, it has been clear that the mRNA sequences represent only a portion of the primary transcripts, and hence specific processing appears to be an essential feature and potential regulatory event in mRNA biogenesis. In lower eukaryotes (i.e., fungi) there is little or no difference in either the complexity or the size of the RNAs found in the nucleus and the cytoplasm (6, 7); in these systems the relationship between nuclear transcripts and translated mRNAs appears to be direct. In higher eukaryotes, where nuclear RNA is several times more complex than mRNA, the relationship between the nuclear and mRNA species is not as clearly understood. In mouse and rat brain (8, 9), RNA-DNA hybridization experiments clearly show that the polyadenylylated nuclear RNA population contains all of the polyadenylylated cytoplasmic RNA sequences. However, it is also clear that both the nuclear RNA and mRNA are composed of poly(A)+-and poly(A)-RNA populations (8)(9)(10). At present, the relationship between the nonadenylylated nuclear RNA and mRNA in these systems is unknown.In Drosophila, however, a recent observation by Lengyel et al. (11) clearly indicates that the transcribed region ofone ofthe major Drosophila heat shock puff sites, 93D, is represented almost exclusively by poly(A)-RNA in the cytoplasm, whereas it is represented by both poly(A)+-and poly(A)-RNA in the nucleus.In view ofthis fact, and because we have determined that the mRNA of Drosophila third-instar larvae is composed of both adenylylated and nonadenylylated species (12), we have measured the sequence complexity of poly(A)+-and total nuclear RNA present in these larvae. We report here that, as was observed in larval mRNA, the nuclear RNA is composed of both an adenyly...

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Section: Resultsmentioning

confidence: 65%

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

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Nonadenylylated mRNA is present as polyadenylylated RNA in nuclei of Drosophila.

Zimmerman¹,

Fouts²,

Lévy³

et al. 1982

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

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“…The mixture was applied to a 1.5~ml protein A-Sepharose C1-4B (Pharmacia) column at 4" C and washed with 80 ml of polyribosome buffer; mRNA in bound polyribosomes was eluted with 3.0 ml of 25 mM Tris, pH 7.6,20 mM EDTA, 20 u/ml of heparin (Shapiro and Young, 1981). RNA in the unbound polyribosome fraction (i.e., depleted polyribosomes) and in the specifically bound polyribosomes (i.e., enriched polyribosomes) was further purified by phenol-chloroform extraction, and polyadenylated RNA was isolated by oligo(dT)-cellulose chromatography (Levy and Manning, 1981). This procedure was repeated until approx.…”

Section: (B) Enrichment Of G6pd Mrnamentioning

confidence: 99%

“…All restriction enzymes were purchased from BRL and used as recommended. Isolation and electrophoresis of nucleic acids and Southern and Northern transfers are described elsewhere (Levy and Manning, 1981;Levy et al, 1982). Hybridization of 5'-end-labeled RNA to Southern blots and filters for library screening was according to Fouts et al (1981).…”

Section: (C) Determination Of Transcription Orientationmentioning

confidence: 99%

Isolation and characterization of the glucose-6-phosphate dehydrogenase gene of Drosophila melanogaster

Ganguly

Manning

1985

Gene

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Polytene chromosomes: The status of the band–interband question

Hill

Rudkin²

1987

BioEssays

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SummaryCracks in the one-gene, one-band paradigm for polytene chromosome organization are widening. A t the same time evidence is accumulating suggesting that decondensed regions of the chromosomes (puffs, diffuse bands, interbands and possibly vacuoles within some bands) are generally associated with gene transcription. A model, now on the ascendancy, is based on the proposal that the bandinterbandpattern is primarily a reflection of local transcriptional state, rather than the distribution of genic and non-genic material.

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Messenger RNA sequence complexity and homology in developmental stages of Drosophila

Cited by 40 publications

References 22 publications

Nonadenylylated mRNA is present as polyadenylylated RNA in nuclei of Drosophila.

Nonadenylylated mRNA is present as polyadenylylated RNA in nuclei of Drosophila.

Isolation and characterization of the glucose-6-phosphate dehydrogenase gene of Drosophila melanogaster

Polytene chromosomes: The status of the band–interband question

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