An improved method for the preparation of undegraded polysomes and active messenger RNA from immature chicken erythrocytes

Perucho, Manuel; Molgaard, H. V.; Shevack, A.; Pataryas, Theocharis A.; Ruiz-Carrillo, Adolfo

doi:10.1016/0003-2697(79)90168-4

Cited by 18 publications

(11 citation statements)

References 20 publications

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“…6B and C). Since H5 mRNA is polyadenylated (29) and the H5 mRNA from drug-treated cells can be translated in vitro (32), the reduction in size was probably due to a shortening of the poly(A) tail. Interestingly, an equivalent effect was not seen with puromycin ( Fig.…”

Section: Resultsmentioning

confidence: 99%

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Regulation of histone and beta A-globin gene expression during differentiation of chicken erythroid cells.

Affolter¹,

Côté²,

Renaud³

et al. 1987

Mol. Cell. Biol.

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The expression of the genes for several histones and pA-globin was examined in the chicken erythroid cell lineage. During the transition from CFU-(E) to the mature erythrocyte, histone H5 gradually increased fourfold in nuclei with little concomitant displacement of the Hi histones. This resulted in a 70% net increase in linker histone (Hi plus H5) content. The differential accumulation of H5 reflected (i) an increase in the transcriptional activity of the H5 gene occurring at the erythroblast stage, (ii) an apparent longer half-life of H5 mRNA, and (iii) a higher stability of the protein. Although the transcriptional activity of the histone genes (except H5) decreased with cell age, it was not tightly coupled to the S phase. On the other hand, the mRNA levels for these histones were tightly regulated during the cell cycle. Use of protein and DNA synthesis inhibitors indicated that the content of H5 mRNA was regulated at the posttranscriptional level by a control mechanism(s) differing from those for the other histones. Although the transcription rates of the H5 and pA_globin genes were comparable, differential accumulation of iA_globin mRNA led to a 30-to 170-fold-higher copy number of the pA-globin mRNA as the cell matured.

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Section: Resultsmentioning

confidence: 99%

“…Mature hen erythrocytes (ME) and immature erythrocytes from peripheral blood (IE-B) were obtained and purified by filtration through SP-Sephadex as described elsewhere (32). Erythroid cells at earlier stages of maturation were obtained from anemic bone marrow.…”

Section: Methodsmentioning

confidence: 99%

Regulation of histone and beta A-globin gene expression during differentiation of chicken erythroid cells.

Affolter¹,

Côté²,

Renaud³

et al. 1987

Mol. Cell. Biol.

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“…Immature hen erythrocytes were obtained and purified as described (15). AEV (RAV-2) transformed chicken erythroblast precursor cells (5) were kindly provided by Dr. T.Graf.…”

Section: Cellsmentioning

confidence: 99%

“…Immature hen erythrocyte polysomes were obtained as described (15). Poly(A) polysomal RNA was prepared by poly(U)-Sepharose (Pharmacia) chromatography (17).…”

Section: Rnamentioning

confidence: 99%

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Construction of chimeric plasmids containing histone H5 cDNA from hen erythrocyte. DNA sequence of a fragment derived from the 5′ region of H5 mRNA

Ruiz‐Vázquez

Ruiz-Carrillo

1982

Nucl Acids Res

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We report the construction and characterization of chicken erythrocyte histone H5 cDNA recombinant plasmids. cDNA was synthesized from poly(A)+ polysomal RNA enriched in H5 mRNA and inserted into the PseI site of pBR322. Several clones containing H5 cDNA sequences were obtained and one of them (p541), expressing H5 antigenic determinants, was sequenced. The DNA insert of p541 contains 118 nucleotides from the 5' non-translated region of H5 mRNA and sequences coding for up to residue 46 of the N-terminus of the arginine (position 15) H5 variant. There is a strikingly high number of repeated sequences both in the leader and coding region; among these, the octanucleotide 5' GCGGCG GC3' is found five times along the sequence. Although the H5 mRNA 5' leader is GC-rich (66%), there is an AT-rich region, about 16 nucleotides long, which shares strong homology with the leaders of sea urchin histone Hi mRNAs.

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Endonuclease G: a (dG)n X (dC)n-specific DNase from higher eukaryotes.

Ruiz-Carrillo¹,

Renaud²

1987

The EMBO Journal

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110

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An endonuclease activity (termed endonuclease G) that selectively cleaves DNA at (dG)". (dC). tracts has been partially purified from immature chicken erythrocyte nuclei. Sites where n 2 9 are cleaved in a manner that resembles types II and HI restriction nucleases. The nicking rate of the Gstrand is 4to 10-fold higher than that of the C-strand depending on the length of the (dG). -(dC). tract and/or nucleotide composition of the flanking sequences. Endonuclease G hydrolyzes (dG)24*(dC)24 of supercoiled DNA in a bimodal way every 9-11 nucleotides, the maxima in one strand corresponding to minima in the opposite, suggesting that it binds preferentially to one side of the double helix. The nuclease produces 5' phosphomonoester ends and its activity is dependent on Mg2+ or Mn2+. The wide distribution and high relative activity of endonuclease G in a variety of tissues and species argues for a general role of the enzyme. The striking correlation between genetic instability and poly(dG) -poly(dC) tracts in DNA suggests that these sequences and endonuclease G are involved in recombination processes.

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An improved method for the preparation of undegraded polysomes and active messenger RNA from immature chicken erythrocytes

Cited by 18 publications

References 20 publications

Regulation of histone and beta A-globin gene expression during differentiation of chicken erythroid cells.

Regulation of histone and beta A-globin gene expression during differentiation of chicken erythroid cells.

Construction of chimeric plasmids containing histone H5 cDNA from hen erythrocyte. DNA sequence of a fragment derived from the 5′ region of H5 mRNA

Endonuclease G: a (dG)n X (dC)n-specific DNase from higher eukaryotes.

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