HeLa cell deoxyribonucleic acid dependent RNA polymerases: function and properties of the class III enzymes

Biochemistry ◽  
1976 ◽  
Vol 15 (7) ◽  
pp. 1500-1509 ◽  
Author(s):  
P. Anthony Weil ◽  
Stanley P. Blatti
Keyword(s):  
Hela Cell ◽  
Deoxyribonucleic Acid ◽  
Rna Polymerases ◽  
Class Iii

scholarly journals HeLa Cell R-Deoxyribonucleic Acid Polymerases

1974 ◽  
Vol 249 (18) ◽  
pp. 5809-5815
Author(s):  
Silvio Spadari ◽  
Arthur Weissbach
Keyword(s):  
Hela Cell ◽  
Deoxyribonucleic Acid

Competitive and cooperative functioning of the anterior and posterior promoter elements of an Alu family repeat

1986 ◽  
Vol 6 (6) ◽  
pp. 2041-2052
Author(s):  
C Perez-Stable ◽  
C K Shen
Keyword(s):  
Hela Cell ◽  
Rna Polymerase Iii ◽  
Trna Genes ◽  
Promoter Element ◽  
Class Iii ◽  
Promoter Elements ◽  
Transcriptional Initiation ◽  
Cell Extracts ◽  
Alu Repeat ◽  
Factor C

Similar to tRNA genes and the VAI gene, the Alu family repeats are transcribed by RNA polymerase III and contain a split intragenic promoter. Results of our previous studies have shown that when the anterior, box A-containing promoter element (5'-Pu-Pu-Py-N-N-Pu-Pu-Py-G-G-3' in which Pu is any purine, Py is any pyrimidine, and N is any nucleotide) of a human Alu family repeat is deleted, the remaining box B-containing promoter element (5'-G-A/T-T-C-Pu-A-N-N-C-3') is still capable of directing weak transcriptional initiation at approximately 70 base pairs (bp) upstream from the box B sequence. This is different from the tRNA genes in which the box A-containing promoter element plays the major role in the positioning of the transcriptional initiation site(s). To account for this difference, we first carried out competition experiments in which we show that the posterior element of the Alu repeat competes with the VAI gene effectively for the transcription factor C in HeLa cell extracts. We then constructed a series of contraction and expansion mutants of the Alu repeat promoter in which the spacing between boxes A and B was systematically varied by molecular cloning. In vitro transcription of these clones in HeLa cell extracts was analyzed by RNA gel electrophoresis and primer extension mapping. We show that when the box A and box B promoter sequences are separated by 47 to 298 bp, the transcriptional initiation sites remain 4 to 5 bp upstream from box A. However, this positioning function by the box A-containing promoter element was lost when the spacing was shortened to only 26 bp or increased to longer than 600 bp. Instead, transcriptional initiation occurred approximately 70 bp upstream from box B, similar to that in the clones containing only the box B promoter element. All the mutant clones were transcribed less efficiently than was the wild type. An increase in the distance between boxes A and B also activated a second box A-like element within the Alu family repeat. We compare these results with the results of tRNA gene studies. We also discuss this comparison in terms of the positioning function of the split class III promoter elements and the evolutionary conservation of the spacing between the two promoter elements for optimum transcriptional efficiency.

scholarly journals Rsc4 Connects the Chromatin Remodeler RSC to RNA Polymerases

2006 ◽  
Vol 26 (13) ◽  
pp. 4920-4933 ◽  
Author(s):  
Julie Soutourina ◽  
Véronique Bordas-Le Floch ◽  
Gabrielle Gendrel ◽  
Amando Flores ◽  
Cécile Ducrot ◽  
...  
Keyword(s):  
Chromatin Remodeling ◽  
Chromatin Accessibility ◽  
Rna Polymerases ◽  
Yeast Genome ◽  
Class Iii ◽  
Promoter Regions ◽  
C Terminus ◽  
Rsc Complex ◽  
The Stability ◽  
Pol Iii

ABSTRACT RSC is an essential, multisubunit chromatin remodeling complex. We show here that the Rsc4 subunit of RSC interacted via its C terminus with Rpb5, a conserved subunit shared by all three nuclear RNA polymerases (Pol). Furthermore, the RSC complex coimmunoprecipitated with all three RNA polymerases. Mutations in the C terminus of Rsc4 conferred a thermosensitive phenotype and the loss of interaction with Rpb5. Certain thermosensitive rpb5 mutations were lethal in combination with an rsc4 mutation, supporting the physiological significance of the interaction. Pol II transcription of ca. 12% of the yeast genome was increased or decreased twofold or more in a rsc4 C-terminal mutant. The transcription of the Pol III-transcribed genes SNR6 and RPR1 was also reduced, in agreement with the observed localization of RSC near many class III genes. Rsc4 C-terminal mutations did not alter the stability or assembly of the RSC complex, suggesting an impact on Rsc4 function. Strikingly, a C-terminal mutation of Rsc4 did not impair RSC recruitment to the RSC-responsive genes DUT1 and SMX3 but rather changed the chromatin accessibility of DNases to their promoter regions, suggesting that the altered transcription of DUT1 and SMX3 was the consequence of altered chromatin remodeling.

scholarly journals ABSENCE OF TRANSLATIONAL CONTROL OF HISTONE SYNTHESIS DURING THE HELA CELL LIFE CYCLE

1970 ◽  
Vol 45 (3) ◽  
pp. 509-513 ◽  
Author(s):  
Thoru Pederson ◽  
Elliott Robbins
Keyword(s):  
Life Cycle ◽  
Hela Cell ◽  
Cytosine Arabinoside ◽  
Translational Control ◽  
Messenger Rna ◽  
Deoxyribonucleic Acid ◽  
S Phase ◽  
Cell Life ◽  
Histone Synthesis ◽  
G1 Cells

The cell-free synthesis of histone-like polypeptides has been achieved using a selected class of small polyribosomes as the only particulate fraction. This synthesis is prevented if the deoxyribonucleic acid (DNA) inhibitor, cytosine arabinoside, is added to the cells prior to disruption, and it is not detected when the cytoplasm used is derived from postmitotic (G1) cells. When the 100,000 g supernate from pure metaphase populations was compared with that from S phase cells, the cell-free synthesis of histone-like polypeptides in the presence of S phase polyribosomes remained unchanged. These data suggest that, except for the histone messenger RNA-ribosome complex, the cytoplasmic factors requisite for histone synthesis are present throughout the cycle, and that the shut-off of this synthesis is not under translational control.

scholarly journals A STUDY OF THE PENETRATION OF MAMMALIAN CELLS BY DEOXYRIBONUCLEIC ACIDS

1961 ◽  
Vol 9 (1) ◽  
pp. 81-91 ◽  
Author(s):  
Ellen Borenfreund ◽  
Aaron Bendich
Keyword(s):  
Nucleic Acids ◽  
Hela Cell ◽  
Perchloric Acid ◽  
Hela Cells ◽  
Mammalian Cells ◽  
Deoxyribonucleic Acid ◽  
Degradation Products ◽  
Dna Bases ◽  
Human Dna ◽  
Nuclear Regions

Tritium-labeled deoxyribonucleic acid (DNA) from pneumococci and from human leukocytes was added to growing cultures of HeLa cells at 37°C. Autoradiography revealed an extensive localization of tritium in the nuclear regions. The label could not be removed by treatment with ribonuclease or dilute perchloric acid, but quantitative removal from the cells could be effected with deoxyribonuclease. Chemical and radioactivity determinations on nucleic acids isolated from the exposed HeLa cells revealed the presence of tritium in all 4 DNA bases. About 12 µg. of tritiated DNA was recovered from 6 x 106 HeLa cells which had been exposed for 24 hours to 240 µg. of the human DNA. From this, it is concluded that the amount of DNA, or its degradation products, taken up by the cells was equivalent to at least 10 per cent of the normal HeLa cell complement.

New deoxyribonucleic acid dependent deoxyribonucleic acid polymerase from HeLa cell mitochondria

Biochemistry ◽  
1973 ◽  
Vol 12 (19) ◽  
pp. 3602-3608 ◽  
Author(s):  
Michael Fry ◽  
Arthur Weissbach
Keyword(s):  
Hela Cell ◽  
Deoxyribonucleic Acid

scholarly journals Competitive and cooperative functioning of the anterior and posterior promoter elements of an Alu family repeat.

1986 ◽  
Vol 6 (6) ◽  
pp. 2041-2052 ◽  
Author(s):  
C Perez-Stable ◽  
C K Shen
Keyword(s):  
Hela Cell ◽  
Rna Polymerase Iii ◽  
Trna Genes ◽  
Promoter Element ◽  
Class Iii ◽  
Promoter Elements ◽  
Transcriptional Initiation ◽  
Cell Extracts ◽  
Alu Repeat ◽  
Factor C

Similar to tRNA genes and the VAI gene, the Alu family repeats are transcribed by RNA polymerase III and contain a split intragenic promoter. Results of our previous studies have shown that when the anterior, box A-containing promoter element (5'-Pu-Pu-Py-N-N-Pu-Pu-Py-G-G-3' in which Pu is any purine, Py is any pyrimidine, and N is any nucleotide) of a human Alu family repeat is deleted, the remaining box B-containing promoter element (5'-G-A/T-T-C-Pu-A-N-N-C-3') is still capable of directing weak transcriptional initiation at approximately 70 base pairs (bp) upstream from the box B sequence. This is different from the tRNA genes in which the box A-containing promoter element plays the major role in the positioning of the transcriptional initiation site(s). To account for this difference, we first carried out competition experiments in which we show that the posterior element of the Alu repeat competes with the VAI gene effectively for the transcription factor C in HeLa cell extracts. We then constructed a series of contraction and expansion mutants of the Alu repeat promoter in which the spacing between boxes A and B was systematically varied by molecular cloning. In vitro transcription of these clones in HeLa cell extracts was analyzed by RNA gel electrophoresis and primer extension mapping. We show that when the box A and box B promoter sequences are separated by 47 to 298 bp, the transcriptional initiation sites remain 4 to 5 bp upstream from box A. However, this positioning function by the box A-containing promoter element was lost when the spacing was shortened to only 26 bp or increased to longer than 600 bp. Instead, transcriptional initiation occurred approximately 70 bp upstream from box B, similar to that in the clones containing only the box B promoter element. All the mutant clones were transcribed less efficiently than was the wild type. An increase in the distance between boxes A and B also activated a second box A-like element within the Alu family repeat. We compare these results with the results of tRNA gene studies. We also discuss this comparison in terms of the positioning function of the split class III promoter elements and the evolutionary conservation of the spacing between the two promoter elements for optimum transcriptional efficiency.

scholarly journals Products of RNA Polymerases in HeLa Cell Nuclei

1971 ◽  
Vol 68 (11) ◽  
pp. 2861-2865 ◽  
Author(s):  
E. A. Zylber ◽  
S. Penman
Keyword(s):  
Hela Cell ◽  
Rna Polymerases ◽  
Cell Nuclei
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