Three abundance classes in HeLa cell messenger RNA

Bishop, John O.; Morton, J.G.; Rosbash, Michael; Richardson, Melville

doi:10.1038/250199a0

Cited by 547 publications

(279 citation statements)

References 42 publications

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“…Analysis of the data obtained as best fit suggests a minimum of three kinetic components. As previously described [21], this result indicates a mRNA population composed of three abundance classes. The percentage of [3 H]cDNA in each abundance class and the corresponding Rot ½ value for that class are presented in Table III.…”

Section: Polysomal Poly(a)*-mrna-[3h]cdna Hybridization the Experimesupporting

confidence: 83%

Complexity and content of the DNA and RNA in Trypanosoma cruzi

Lanar

Lévy

Manning

1981

Molecular and Biochemical Parasitology

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The content and sequence complexity of the nuclear DNA and messenger RNA for epimastigotes of Trypanosoma cruzi were determined. From analysis of nuclear DNA reassociation studies and microspectrofluorometric measurements of laser induced fluorescence of cellular DNA, T. cruzi is found to be a diploid organism with a nuclear DNA content of 2.5 x 108 nucleotide pairs (2.8 X 10 -~3 g) and a kinetoplast DNA content of 4.9 X 107 nucleotide pairs (5.4 X 10 -t4 g). Reassoeiation kinetics of nuclear DNA of average length 0.4 kb reveals three kinetic components: a moderately repetitive component with a reiteration frequency of 5.1 X 103 present in 9% of the fragments, a lowly repetitive component with a reiteration frequency of 32 present in 51% of the fragments, and a single-copy component present in 23% of the fragments.By saturation hybridization of total polysomal RNA to 3H-labeled single-copy DNA, it was determined that 68% of the single-copy DNA was represented in the epimastigote polysomal RNA. This corresponds to ca. 12 000 different mRNA species. Of these, ca. 9000 are present as poly(A)÷-RNA, while the remaining 3000 appear not to be polyadenylated. Kinetic analysis of the poly(A)+-RNA population indicates it is composed of at least three classes of RNAs of different abundancy levels: two sequences which occur ca. 3000 times per cell, ca. 750 sequences which occur about 20 times per cell, and ca. 15 500 sequences which occur 1-2 times per cell.

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Section: Polysomal Poly(a)*-mrna-[3h]cdna Hybridization the Experimesupporting

confidence: 83%

Complexity and content of the DNA and RNA in Trypanosoma cruzi

Lanar

Lévy

Manning

1981

Molecular and Biochemical Parasitology

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“…2A), indicates that the cDNA represents the diversity of its template mRNA population. Additional arguments supporting this have also been given by others [30,31,14,24].…”

Section: Isolation Of Mrna and Reverse Transcription Into Complementamentioning

confidence: 69%

“…This is due to the fact that the mRNA species occur in widely varying frequencies, whereby the more abundant mRNAs hybridize at lower rot values than the more scarce mRNAs. Similar to the hybridization kinetics for other mRNA cell populations [30,31,36,241 the complex curve can be divided into 3 kinetic components, representing 3 mRNA frequency classes [30]. Table 2 shows the numerical evaluation of the hybridization data plotted in Fig.…”

Section: Cellular Amounr Andpo!y(a) Content Of Polv(a)-contuining mentioning

confidence: 99%

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Frequency Distribution of Messenger Sequences within Polysomal mRNA and Nuclear RNA from Rat Liver

Sippel

Hynes

Groner

et al. 1977

European Journal of Biochemistry

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DNA complementary to polysomal poly(A)-containing mRNA (cDNA) of male rat liver was used to study the diversity of messenger sequences in the nucleus and in polysomes.1. Hybridization of cDNA against an excess of its own polysomal mRNA template revealed that about 10000 different mRNA species are expressed in the liver tissue. They are distributed in a wide frequency range derived from approximately 0.5 % of the total genome.2. Hybridization of the cDNA against total nuclear RNA shows that messenger sequences comprise less than 1 of the mass of total nuclear RNA. Messenger sequences have a different frequency distribution in nucleus and cytoplasm.3. In hybridizations using cDNA, which had been fractionated into sequences representing abundant and scarce polysomal mRNA molecules, it was found that although abundant cytoplasmic messenger sequences are also abundant in the nucleus, they exist in a significantly lower frequency range in the nuclear compartment.Different rates of synthesis of a specific protein are mediated by parallel changes of the functional level of the corresponding mRNA in a variety of systems [l -51. Mechanisms responsible for altering mRNA concentrations are not well understood, since our knowledge of nuclear processes in the biogenesis of eucaryotic mRNA is limited. Molecular hybridization techniques have brought some insight into the relationship between nuclear and cytoplasmic RNA. It has recently been shown that nuclear RNA is up to 10 times as complex as polysomal mRNA [6-121. Poly(A)-containing nuclear RNA, generally assumed to be the RNA pool from which most cytoplasmic mRNA molecules are derived (for review see [13]), was found to have a significantly higher complexity than polysomal mRNA in a variety of cell types [14-161. From the observed reductions in RNA sequence complexity and RNA size (see accompanying paper), it is clear that only a minor subset of sequences in Abbreviations. cDNA, DNA copy complementary to mRNA; hnRNA, polydisperse high-molecular-weight nuclear RNA ; AMV, avian myeloblastosis virus. rot value, concentration of RNA (as mol phosphate/l) times time of hybridization (in s); rof1p, rot at which half of hybridizable cDNA is hybridized.Enzymes. Staphylococcal nuclease, nucleate 3'-oligonucleotidohydrolase (EC 3.1.4.7) ; reverse transcriptase, RNA-dependent DNA polymerase, deoxynucleosidetriphosphate: DNA deoxynucleotidyltransferase (EC 2.7.7.7); DNase I, deoxyribonucleate oligonucleotidohydrolase (EC 3.1.4.5).nuclear RNA leaves the nucleus to become active polysomal mRNA. Nuclear polyadenylation is certainly not a sufficient prerequisite for an RNA sequence to be selected for transport into the cytoplasm.Very little information exists on the frequency distribution of messenger sequences within total nuclear RNA. To obtain more information we have performed a comparative examination of the kinetics of hybridization of nuclear RNA and polysoma1 mRNA to complementary DNA synthesized from poly(A)-containing mRNA. We have found that a different frequency distribution of ...

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“…It usually shows a pattern of multiple transitions which suggests that the frequencies of individual messenger species are clustered around 2 to 4 discrete values, rather than taking any value in a broad, continuous range. In other words, the quantitative structure of an mRNA population could be approximately described by dividing it into 2 to 4 abundance classes (2)(3)(4).…”

Section: Introductionmentioning

confidence: 99%

Frequency distribution of mRNA and pre-mRNA in growing and differentiated Friend cells

Mauron¹,

Spohr²

1978

Nucl Acids Res

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The frequency distribution of poly(A)+-mRNA in growing and in differentiated Friend cells has been measured by mRNA-cDNA hybridization and their differences established by heterologous hybridization of mRNA of one type and cDNA of the other. It was shown that induction of Friend cells involves an increase in abundance of a small number of mRNAs, while no specific pattern of messenger disappearance could be detected. The frequency distribution of pre-mRNA was determined by hybridizing nuclear RNA with the cDNA probes complementary to mRNA. In uninduced Friend cells, it was shown that most precursor messenger sequences are present at a single frequency of about 3 molecules per nucleus, independently of their final frequency in polysomal mRNA. In induced Friend cells, the frequency distribution of premRNA is more heterogeneous and correlated to some extent with the corresponding mRNA frequency distribution. INTRODUCTIONThe control of gene expression in eukaryotic cells still defies a general explanation, partly because of the multiplicity of levels at which molecular regulation devices may operate. In order to appreciate the importance of these various levels of control, it would be important to know which sets of genes are transcribed into nuclear RNA and into messenger RNA in a given cell type.

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Three abundance classes in HeLa cell messenger RNA

Cited by 547 publications

References 42 publications

Complexity and content of the DNA and RNA in Trypanosoma cruzi

Complexity and content of the DNA and RNA in Trypanosoma cruzi

Frequency Distribution of Messenger Sequences within Polysomal mRNA and Nuclear RNA from Rat Liver

Frequency distribution of mRNA and pre-mRNA in growing and differentiated Friend cells

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