The core promoter of mouse rDNA consists of two functionally distinct domains

Clos, Joachim; Normann, Andrea; Öhrlein, Andrea; Grummt, Ingrid

doi:10.1093/nar/14.19.7581

Cited by 46 publications

(30 citation statements)

References 21 publications

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“…The conserved guanine residue at nucleotide -16 has an important role in this binding, as shown by previous studies (15,30,33; Tanaka et al, unpublished observation). On the other hand, conserved nucleotides -1 and -7 are presumably crucial for the relatively speciesindependent recognition processes of rDNA such as the binding and functioning of polymerase I enzyme or other factor(s) (3,14,30). The role of upstream sequences was not clarified in this study, except that some protein in the TFID fraction bound with the region of nucleotides -40 to -140 in a core sequence-dependent manner.…”

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confidence: 75%

“…The former promoter element lies proximal to the transcription start site and is called a core promoter. For the mouse rDNA, this region was shown to be around nucleotides -40 to +23 by deletion experiments (3,25,41). Further upstream sequences are required for higher transcription efficiency, which is apparently revealed under certain stringent conditions in vitro (26).…”

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confidence: 98%

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Structural determinant of the species-specific transcription of the mouse rRNA gene promoter.

Sáfrány¹,

Tanaka²,

Kishimoto³

et al. 1989

Mol. Cell. Biol.

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Mammalian ribosomal DNA (rDNA) transcription has a certain species specificity such that, both in vivo and in vitro, human rDNA cannot be transcribed by mouse machinery and vice versa. This is due to a species-dependent transcription factor, TFID (Y. Mishima, I. Financsek, R. Kominami, and M. Muramatsu, Nucleic Acids Res. 10:6659-6670, 1982). On the basis of the information obtained from 5' and 3' substitution mutants, we prepared a chimeric gene in which the mouse sequence from positions -32 to -14 was inserted into the corresponding location of the human rDNA promoter. The chimeric gene could be transcribed by mouse extracts nearly as efficiently as the wild-type mouse promoter. The chimeric gene could also sequester transcription factor TFID at an efficiency similar to that for the mouse promoter. Partially purified mouse TFID that could not protect the human rDNA promoter against DNase I produced a clear footprint on this chimeric gene that was similar to that on mouse rDNA promoter. The basic structure of the mouse rDNA core promoter is discussed in relation to the interaction with TFID.

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confidence: 75%

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confidence: 98%

Structural determinant of the species-specific transcription of the mouse rRNA gene promoter.

Sáfrány¹,

Tanaka²,

Kishimoto³

et al. 1989

Mol. Cell. Biol.

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“…DNase I footprinting experiments with mSL1 map a protected region between -1 2 and -5 2 of the promoter {Fig. 3, lane 4), sequences previously shown to be important for the formation of stable transcription complexes at the mouse promoter (Clos et al 1986b;Tower et al 1986;Nagamine et al 1987). D N A binding experiments have also been performed by use of two point mutations at nucleotides -7 and -1 6 of the mouse core element {the generous gift of M. Muramatsu).…”

Section: Msl1 and M Ubf Form A Cooperative Dna-binding Complexmentioning

confidence: 99%

Assembly of alternative multiprotein complexes directs rRNA promoter selectivity.

Bell¹,

Jantzen²,

Tjian³

1990

Genes Dev.

159

122

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How can trans-activators with the same DNA binding specificity direct different transcriptional programs? The rRNA transcriptional apparatus offers a useful model system to address this question and to dissect the mechanisms that generate alternative transcription complexes. Here, we compare the mouse and human transcription factors that govern species-specific RNA polymerase I promoter recognition. We find that both human and mouse rRNA transcription is mediated by a specific multiprotein complex. One component of this complex is the DNA-binding transcription factor, UBF. Paradoxically, human and mouse UBF display identical DNA binding specificities even though transcription of rRNA is species specific. Promoter selectivity is conferred by a second essential factor, SL1, which, for humans, does not bind DNA independently but, instead, cooperates with UBF in the formation of high-affinity DNA-binding complexes. In contrast, mouse SLI can selectively interact with DNA in the absence of UBF. Reconstituted transcription experiments establish that UBF and RNA polymerase I from the two species are functionally interchangeable, whereas mouse and human SL1 exhibit distinct DNA binding and transcription activities. Together, these results suggest a critical role for a specific multiprotein assembly in RNA polymerase I promoter recognition and reveal distinct mechanisms through which such complexes can generate functional diversity.[Key Words: RNA pol I; rRNA transcription; species specificity; transcription factors; UBF; SL1] Received February 7, 1990; revised version accepted March 14, 1990.Transcriptional initiation in eukaryotic cells is a highly regulated process requiring the correct association of numerous proteins into a specific complex with RNA polymerase {Mitchell and Tjian 1989; Saltzman and Weinmann 1989). Although mapping of essential promoter elements has identified multiple DNA sequences important for template recognition, how the different transcription factors work together with RNA polymerase to select these sequences as sites of initiation is poorly understood. Current results suggest strongly that the DNA sequence specificity of a single protein cannot account for promoter recognition by any of the three cellular RNA polymerases {Yoshinaga et al. 1987; Murphy et al. 1989; Smale and Baltimore 19891.Instead, it seems that the interactions of multiple proteins, both with the DNA and with one another, are required to generate the observed selectivity of initiation.The specificity of RNA polymerase I (RNA pol I} transcription is well suited for studies of promoter recognition. Unlike RNA pol II and RNA pol III, which recognize a wide variety of promoters, RNA pol I apparently initiates from only a single type of promoter in the cell, that of the large rRNA gene . This limited promoter range is reflected in the stringent species specificity of RNA pol I transcription.Studies using either intact cells or in vitro transcription extracts indicate that whereas very closely related species [e.g., mouse and rat)...

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“…The major difference was that the 5Ј⌬-33 construct was expressed at approximately the same level as the Ϫ55 construct in vitro whereas in vivo, the Ϫ33 construct was only weakly expressed, at best. A parallel is that in mammalian systems, core promoter sequences are often sufficient to program transcription in vitro, but additional upstream sequences are needed for efficient transcription in vivo (35)(36)(37)(38).…”

Section: Development Of a Plant In Vitromentioning

confidence: 99%

Extensive purification of a putative RNA polymerase I holoenzyme from plants that accurately initiates rRNA gene transcription in vitro

Sáez‐Vasquez

Pikaard

1997

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

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RNA polymerase I (pol I) is a nuclear enzyme whose function is to transcribe the duplicated genes encoding the precursor of the three largest ribosomal RNAs. We report a cell-free system from broccoli (Brassica oleracea) inf lorescence that supports promoter-dependent RNA pol I transcription in vitro. The transcription system was purified extensively by DEAE-Sepharose, Biorex 70, Sephacryl S300, and Mono Q chromatography. Activities required for pre-rRNA transcription copurified with the polymerase on all four columns, suggesting their association as a complex. Purified fractions programmed transcription initiation from the in vivo start site and utilized the same core promoter sequences required in vivo. The complex was not dissociated in 800 mM KCl and had a molecular mass of nearly 2 MDa based on gel filtration chromatography. The most highly purified fractions contain Ϸ30 polypeptides, two of which were identified immunologically as RNA polymerase subunits. These data suggest that the occurrence of a holoenzyme complex is probably not unique to the pol II system but may be a general feature of eukaryotic nuclear polymerases.Eukaryotic nuclear RNA polymerases interact with transcription factors to recognize gene promoters and initiate transcription. Many of the required factors can be purified individually and added back together to reconstitute transcription in vitro, allowing their identification and molecular characterization (1-7). However, proteins not required under a given set of assay conditions can be overlooked, though they may play important roles in vivo. The latter insight has come, in part, from the characterization of RNA pol II holoenzyme complexes in yeast and mammals (8)(9)(10)(11)(12). These complexes are several megadaltons in size and include pol II core enzyme subunits, most of the general transcription factors, and a growing list of additional activities including protein kinases (13), chromatin remodeling activities(14), and proteins involved in DNA repair (12).There has been at least one report of a pol I-containing fraction from rat cell-free extracts sufficient for accurate transcription in vitro (15), and several studies have shown that pol I transcription factors interact. For instance, the vertebrate pol I transcription factors UBF and SL1͞TIF-IB interact both in solution and on the DNA (16-18). Likewise, UBF and pol I cosediment in glycerol gradients and interact on Far-Western blots (19). Two additional factors, TIF-IA and TIF-IC copurify with pol I on multiple columns (20,21). Collectively, these results suggest that the pol I transcription factors might be associated with one another, possibly within a complex analogous to the pol II holoenzyme.Encouraged by brief reports that cell-free rRNA gene transcription could be achieved in plant extracts (22, 23), we set out to develop a cell-free system from broccoli (Brassica oleracea) to further our studies of rRNA gene regulation in Brassica and Arabidopsis (24-28). We show that a pol Icontaining activity sufficient for pr...

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The core promoter of mouse rDNA consists of two functionally distinct domains

Abstract: SUMMARYWe have determined the sequences constituting the minimal promoter of mouse rDNA. A very small region immediately upstream of the transcription start site (from -1 to -39) is sufficient to direct correct transcription initiation.

Cited by 46 publications

References 21 publications

Structural determinant of the species-specific transcription of the mouse rRNA gene promoter.

Structural determinant of the species-specific transcription of the mouse rRNA gene promoter.

Assembly of alternative multiprotein complexes directs rRNA promoter selectivity.

Extensive purification of a putative RNA polymerase I holoenzyme from plants that accurately initiates rRNA gene transcription in vitro

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