Transcription of Polydeoxythymidylate Sequences in the Genome of the Cellular Slime Mold,
            <i>Dictyostelium discoideum</i>

Jacobson, Allan; Firtel, Richard A.; Lodish, Harvey F.

doi:10.1073/pnas.71.5.1607

Cited by 39 publications

(22 citation statements)

References 28 publications

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“…Since previous work indicated that about half of the DNA is transcribed into mRNA (7), it appears that there is, on the average, one poly(dT)25 sequence in the Dictyostelium genome per mRNA encoded. Additional hybridization experiments showed that the poly(dT) sequences are interspersed throughout the genome, and not clustered in one region (23). Thus, it appears that most DNA sequences coding for mRNA terminate with a poly(dT) sequence, as is indicated schematically in Fig.…”

Section: Following Addition Of [3h]adenosine To Vegetative Cellsmentioning

confidence: 90%

“…The other class is very heterogeneous and contains some sequences migrating at about 6 S [poly(A),5o] which are larger than is found in mRNA, and also material smaller than that isolated from mRNA; possibly the intermediate (2-4S) sized material represents nascent, growing poly(A) chains. Based on calculations similar to those for mRNA, we conclude that each molecule of nuclear poly(A)-containing RNA (isolated from pulse-labeled cells) contains one sequence of poly-(A)25 while only one molecule in three contains a longer poly-(A) sequence (13,23).…”

Section: Synthesis Of Mrnamentioning

confidence: 99%

“…5) (22, 23). Nucleotide analysis of the putative poly-(A)loo and poly(A)25 labeled with 32p showed that they indeed were poly(A) sequences (23 (23). Pulse-labeled nuclear poly(A)-containing RNA also contains two classes of poly(A) sequences (Fig.…”

Section: Synthesis Of Mrnamentioning

confidence: 99%

“…The poly(A)1oo_1ro sequence is added during the final 2-min period in the nucleus, perhaps coincidentally with the loss of the repetitive transcript at the 5' end. Poly(dT)25 sequences in Dictyostelium DNA Dictyostelium DNA contains sufficient poly(dT) sequences of approximate size to encode the poly(A)25 sequences in mRNA (13,23). To demonstrate this, 2P-labeled Dictyostelium DNA was depurinated (24) and poly(dT) sequences were purified by poly(A)-Sepharose chromatography.…”

Section: Following Addition Of [3h]adenosine To Vegetative Cellsmentioning

confidence: 99%

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Synthesis of Messenger RNA and Chromosome Structure in the Cellular Slime Mold

Lodish

Jacobson

Firtel

et al. 1974

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

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Section: Following Addition Of [3h]adenosine To Vegetative Cellsmentioning

confidence: 90%

Section: Synthesis Of Mrnamentioning

confidence: 99%

Section: Synthesis Of Mrnamentioning

confidence: 99%

Section: Following Addition Of [3h]adenosine To Vegetative Cellsmentioning

confidence: 99%

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Synthesis of Messenger RNA and Chromosome Structure in the Cellular Slime Mold

Lodish

Jacobson

Firtel

et al. 1974

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

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“…The length of this sequence varies from 200 nucleotides in mammalian cells [39] to about 150 in insects (Pieris brassicae [40]), 100-120 in plants (Phaseolus aureus [6]) and 50 -60 in lower eukaryotic organism (Saccharomyces cerevisae [4] , Dictyostelium disoideum [41]). We have found an electrophoretic mobility corresponding to 4.5 S for the poly(A) from Tetrahymena pyriformis nuclear and cytoplasmic mRNAs, which corresponds to a chain length of about 100 nucleotides assuming that the electrophoretic behaviour of poly(A) is similar to that of tRNA and 5-S rRNA of E. coli.…”

Section: Discussionmentioning

confidence: 99%

Poly(A)‐Containing RNA in Tetrahymena pyriformis

Rodriguez-Pousada

Hayes

1976

European Journal of Biochemistry

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Poly(A)-containing RNAs with sedimentation coefficients in the range 10-45 S and 10-35 S ( M , 0.2-3.3 x lo6 and 0 . 2 -2 . 2~ lo6 approximately) are present in the nucleus and cytoplasm of T. pyriformis respectively. The poly(A) segments present in both nuclear and cytoplasmic poly(A)-containing RNAs have a chain length of 80-150 nucleotides. Significant amounts of highmolecular-weight poly(A)-containing RNAs (35 s, M , 2.2 x 1 0 6 ) are found in the post-polysomal fraction of the cytoplasm (messenger ribonucleoprotein particles?) and also in large polysomes. In order to exclude the possibility that these high-molecular-weight species are products of nonspecific aggregation, RNA preparations were submitted to severe denaturing conditions before analysis and all electrophoreses were carried out in the presence of 95 % (v/v) formamide.The currently accepted model for mRNA synthesis in eukaryotic cells, proposed by Darnel1 and coworkers [l], involves synthesis of large precursor RNAs in the cell nucleus, post-transcriptional addition of poly(adeny1ic acid) sequences to their 3' termini, reduction of the size of the poly(A)-containing RNAs without loss of the poly(A) segments and transfer of the shortened molecules to the cytoplasm. As required by this model, poly(A) sequences have been found in cytoplasmic and nuclear RNA of mammalian cells [2], and of a variety of lower eukaryotic organisms [ 3 -71. We have previously described several species of nonmethylated cytoplasmic RNA in the unicellular eukaryote, Tetrahymena pyriformis [8], and in subsequent work have found that their RNAs contain poly(A) sequences. In this report we describe how poly(A)-containing RNAs isolated from the nucleus, cytoplasm and polysomes of T. pyriformis have been characterised by electrophoretic analysis under strongly denaturing conditions to avoid aggregation [9,10] and by estimation of the lengths of their poly(A) segments. MATERIALS AND METHODS ChemicalsTriethanolamine (analytical grade) and formamide (technical grade) were supplied by Prolabo, France. The latter was purified by fractional distillation in vucuo followed by treatment with Amberlite MB-1 mixed-bed ion-exchange resin (B.D.H., England) according to Pinder et al. [lo]. Acrylamide (Fluka, Switzerland) and N,N'-bisacrylamide (Eastman, Kodak, U.S.A.) were purified by recrystallisation according to Loening [ll]. Cyanogen bromide was supplied by Eastman Kodak, U.S.A., dextran sulfate and Sepharose 4B by Pharmacia, Sweden ; pancreatic RNase, and RNase A by Sigma, U.S.A.; and T1 RNase by Sankyo, Japan. Other products were obtained from Prolabo, France, or Merck, Germany.[S3H]Uridine (spec. act. 20-28 Ci/mmol), and [2-3H]adenine (spec. act. 15 -25 Ci/mmol) were supplied by the Commissariat a 1'Energie Atomique, France. BuffersBuffer 1 = 10 mM Tris-HC1 pH 7.5,lO mM MgClz, 0.1 M NaC1, 5 pg/ml cycloheximide and 40 pg/ml dextran sulfate. Buffer 2 = 10 mM Tris-HC1 pH 7.6, 0.1 M NaCl, 10 mM EDTA, 0.2% sodium dodecyl sulfate. Buffer 3 = 10 mM Tris-HC1 pH 7.5, 0.36 M NaCl. Buffer 4 = 5...

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