Protein-binding site at the immunoglobulin mu membrane polyadenylylation signal: possible role in transcription termination.

Law, Ronald E.; Kuwabara, Michio D.; Briskin, Michael; Hermanson, Gary; Sigman, David S.; Wall, Randolph

doi:10.1073/pnas.84.24.9160

Cited by 32 publications

(20 citation statements)

References 36 publications

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“…For example, Gilmour and Lis (29) have described protein-DNA cross-linking experiments that show an accumulation of polymerases near the 5' ends of several Drosophila genes, and Chinsky et al (15) have shown, by run-on analysis, that the mouse adenosine deaminase gene exhibits 5' truncation of transcription in tissues in which expression of the gene is low. However, the most fully characterized examples are found among viral transcription units (33,37,41,51,57,74), cellular oncogenes (5,6), and the immunoglobulin jx-b transcription unit (47,53,80).…”

Section: Discussionmentioning

confidence: 99%

Active beta-globin gene transcription occurs in methylated, DNase I-resistant chromatin of nonerythroid chicken cells.

et al. 1990

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The nature and function of the transcriptionally competent state of chromatin remains an enigma despite much work. Two properties are generally characteristic of competent chromatin: an increased sensitivity to digestion by DNase I and a reduced frequency of cytosine methylation at CpG positions (63). However, the notion of "competent" chromatin is poorly defined. Generally it is taken to mean a region of chromatin that is committed, by virtue of a special conformation, to the present or future transcription of one or more of its resident genes.Little is known about the nature of the DNase I-sensitive conformation of competent chromatin. The DNase I-sensitive state encompasses both the active genes themselves and the nontranscribed regions of chromatin flanking them. This contrasts with the situation for transcriptionally induced perturbations of chromatin, for which several structural features have been described (3,14,18,22,40,58,67,79). These perturbations are limited both temporally and spatially to the regions actually undergoing the transcription and are therefore distinct from the all-inclusive DNase I sensitivity of competent chromatin. The large DNase I-sensitive regions characteristic of competent chromatin may include several genes and, in the most completely mapped instances, cover discrete domains that extend uniformly through 10 to 100 kilobase pairs (kb) of chromatin (2, 39, 49). These domains can even include more nontranscribed flanking chromatin than actively transcribed gene chromatin. Moreover, the nontranscribed regions are just as sensitive to DNase I digestion as are the transcribed regions, provided the last-cut approach to the measurement of DNase I sensitivity is used (2,39,49,67,72).

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Section: Discussionmentioning

confidence: 99%

Active beta-globin gene transcription occurs in methylated, DNase I-resistant chromatin of nonerythroid chicken cells.

et al. 1990

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“…RNA polymerase II transcription termination occurs hundreds or thousands of nucleotides downstream of polyadenylation sites (7,13,14). Although there have been a number of reports of specific sequences which may act as termination sequences (15)(16)(17)(18)(19)(20)(21)(22)(23)(24)(25), in vivo, transcription termination often depends on 3'-end processing (9)(10)(11)(12)26). Mutations in the conserved polyadenylation AAUAAA signal of the mouse major 3-globin gene or the human globin gene, respectively (11,12), or mutations in both the AAUAAA hexanucleotide and the downstream GT-rich region of the SV40 early polyadenylation site (9) can inhibit polyadenylation as well as transcription termination.…”

Section: Introductionmentioning

confidence: 99%

Polyadenylation and transcription termination in gene constructs containing multiple tandem polyadenylation signals

1994

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The processes of pre-mRNA 3'-end cleavage and polyadenylation have been closely linked to transcription termination by RNA polymerase II. We have studied the relationship between polyadenylation and transcription termination in gene constructs containing tandem poly(A) signals, at least one of which is the inefficient polyomavirus late poly(A) site. When identical tandem viral signals were separated by fewer than 400 bp, they competed for polyadenylation. The upstream site was always chosen preferentially, but relative site choice was influenced by the distance between the signals. All of these constructs showed the same low level of transcription termination as wild type polyomavirus, which contains a single late poly(A) site. When tandem poly(A) signals were not identical, a stronger downstream signal could outcompete a weaker upstream signal for polyadenylation without altering the efficiency of transcription termination characteristic for use of the upstream signal. Thus, if a weak polyoma virus late poly(A) signal (associated with inefficient transcription termination) preceded a strong rabbit 3-globin signal (associated with efficient transcription termination), termination remained inefficient, but the distal signal was most often chosen for polyadenylation. These results are consistent with independent regulation of polyadenylation and transcription termination in this system and are discussed in light of current models for the dependence of transcription termination on a functional poly(A) site.

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“…Termination is elicited by the binding of a specific termination factor, TTF-1, to this sequence (3,17). In mRNA-encoding genes, it has been suggested that protein binding mediates poly(A) site-dependent termination in transcription of the pu-8 immunoglobulin gene (23). Law et al found that termination occurred in a region located 500 to 1,000 bp downstream from the RUm poly(A) site in myeloma cells, but no detectable termination within this region occurred in pre-B cells (23).…”

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confidence: 99%

“…In mRNA-encoding genes, it has been suggested that protein binding mediates poly(A) site-dependent termination in transcription of the pu-8 immunoglobulin gene (23). Law et al found that termination occurred in a region located 500 to 1,000 bp downstream from the RUm poly(A) site in myeloma cells, but no detectable termination within this region occurred in pre-B cells (23). Gel retention assays revealed DNA-protein complexes of different mobilities when DNA fragments were incubated in nuclear extracts prepared from myeloma and pre-B cells, suggesting that differential protein binding may have a role in the differential termination observed in the two cell types.…”

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RNA polymerase II transcription termination is mediated specifically by protein binding to a CCAAT box sequence.

Connelly¹,

Manley²

1989

Mol. Cell. Biol.

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A region in the adenovirus major late promoter (MLP) containing a CCAAT consensus sequence can direct transcription termination of RNA polymerase II, The mechanisms responsible for transcription termination of mRNA-encoding genes in eucaryotic cells are poorly understood. Several recent studies have shown that efficient termination of transcription by RNA polymerase II (pol II) requires a functional poly(A) addition site (8,22,25,36). However, transcription continues well beyond this site before terminating (for reviews, see references 4 and 31), and another signal that delineates the region or site at which termination occurs is necessary to direct efficient termination (9, 25). We have shown that a sequence containing an inverted CCAAT box consensus in the adenovirus major late promoter (MLP) functions as such a termination site signal (9). Since a number of proteins that bind CCAAT box sequences have been recently identified (6,7,10,20,26,32) and, more specifically, a protein purified from HeLa cells, CP1, has been shown to bind to the CCAAT sequence within the MLP (20), it is possible that protein binding to the MLP CCAAT box mediates termination induced by this sequence. However, the terminator regions analyzed previously all contained MLP sequences in addition to the CCAAT consensus. Furthermore, the ability of the fragments to induce termination was orientation dependent, perhaps suggesting that a function other than protein binding is involved.To gain insights into the mechanism by which this termination site functions, we have investigated the ability of a short oligonucleotide containing either the CCAAT consensus or mutant derivatives to elicit termination when inserted downstream of a poly(A) site. For these studies, p4)4-SVA, a chimeric plasmid that consists of the MLP (Ad2 nucleotides [nt] -405 to +33) directing transcription of the simian virus 40 (SV40) early region (SV40 nt 5171-2533) in a pBR322 vector (Fig. 1A) (24), was used to direct transcription in transient expression assays. The plasmid pSV2.CAT (12) was cotransfected in all cases to standardize transfection efficiencies (8, 9). Assays were performed using human 293 cells (1, 13), which have been shown to support high levels of transcription from p404-SVA (24 from -125 to -73 relative to the transcription start sites, and the other containing sequences from -85 to -51, were both capable of inducing termination when inserted into the SalI site (pBR322 nt 652) of p4)4-SVA located 330 base pairs (bp) downstream from the SV40 poly(A) site (9). These results together suggested that MLP sequences between -85 and -73 may be sufficient to induce termination.To test this hypothesis directly, a 13-bp oligonucleotide containing this sequence was inserted into the Sall site of p4)4-SVA (Fig. 1B). Plasmids containing single insertions in either the orientation in which this sequence is naturally found within the MLP, wt(1+), or in the opposite orientation, wt(1-), were identified and transfected into 293 cells. Forty-eight hours later, nuclei were ...

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Protein-binding site at the immunoglobulin mu membrane polyadenylylation signal: possible role in transcription termination.

Cited by 32 publications

References 36 publications

Active beta-globin gene transcription occurs in methylated, DNase I-resistant chromatin of nonerythroid chicken cells.

Active beta-globin gene transcription occurs in methylated, DNase I-resistant chromatin of nonerythroid chicken cells.

Polyadenylation and transcription termination in gene constructs containing multiple tandem polyadenylation signals

RNA polymerase II transcription termination is mediated specifically by protein binding to a CCAAT box sequence.

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