Stimulation of sea urchin H2B histone gene transcription by a chromatin-associated protein fraction depends on gene sequences downstream of the transcription start site.

Mous, Jan; Stunnenberg, Hendrik G.; Georgiev, Oleg; Birnstiel, Max L.

doi:10.1128/mcb.5.10.2764

Cited by 35 publications

(22 citation statements)

References 27 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Microinjection into oocytes or fertilized eggs of the african clawed toad Xenopus laevis has long been used to study transcription processes (Mous et al, 1985;Gurdon and Wakefield, 1986 and references therein). In the oocyte, there is generally high transcriptional activity, which attenuates as genes are inactivated upon maturation towards the egg.…”

Section: Introductionmentioning

confidence: 99%

Different Potential of Cellular and Viral Activators of Transcription Revealed in Oocytes and Early Embryos ofXenopus laevis

Xu¹,

Rungger²,

Georgiev³

et al. 1994

Biological Chemistry Hoppe-Seyler

View full text Add to dashboard Cite

Many protein domains for transcriptional activation also function when fused to a heterologous DMA binding domain. In mammalian HeLa cells, we have previously characterized the activation domains of several transcription factors using GAL4 fusion proteins. Here we have tested their transcriptional activity in oocytes and developing embryos of the clawed toad Xenopus laevis. We find that the "acidic 11 C-terminal domain of the herpesvirus VP16 (= Vmw65) activator, which is active from yeast to man, is also very active in the two Xenopus systems. The constitutive nature of this viral domain may have evolved to be refractory to cellular defense mechanisms. By contrast, activation domains from cellular eukaryotic transcription factors (TFE3, ITF2, MTF-1) are differentially active in oocytes and early embryos. This indicates that their activity can be regulated by protein modification and/or availability of specific coactivators. We have also compared VP16 induced enhancement of transcription from remote and promoter-proximal positions. In both oocytes and late blastula embryos, activation from a promoter-proximal position was more than 50 fold, while only a moderate stimulation (3-8 fold) was observed from remote positions. This may mean that frog oocyte and early embryos are not yet fully geared for gene control by remote enhancers, i.e. respond predominantly to close-by regulatory sequences. The fact that cellular enhancers are naturally located at various distances from the responsive promoters may thus be exploited by multicellular organisms for differential gene control at early and late stages of development.

show abstract

Section: Introductionmentioning

confidence: 99%

Different Potential of Cellular and Viral Activators of Transcription Revealed in Oocytes and Early Embryos ofXenopus laevis

Xu¹,

Rungger²,

Georgiev³

et al. 1994

Biological Chemistry Hoppe-Seyler

View full text Add to dashboard Cite

show abstract

“…A variety of genes, including histone genes, are accurately transcribed after microinjection into oocyte nuclei, and the injection of genes with mutated promoter regions has enabled the identification of elements required for transcription in the oocyte (16). The Xenopus oocyte system has also been used to identify a trans-acting factor involved in the processing of sea urchin histone mRNAs (34) and, more recently, to identify a factor capable of stimulating sea urchin early H2b gene transcription (30).…”

mentioning

confidence: 99%

“…The TflIIA factor binds with a fourfold-higher affinity to the somatic gene subfamily than to the oocyte gene subfamily. This difference in affinity, together with a decline in the intranuclear concentration of TflIIA during development, has been proposed as an explanation for the inactivation of the oocyte 5S genes in latestage embryos (5,37 Mous et al (30) showed that a 0.3 M salt extract of sea urchin (Psammechinus miliaris) chromatin stimulates expression of early H2b histone genes in Xenopus oocytes. (Late histone genes were not tested.)…”

mentioning

confidence: 99%

Differential stimulation of sea urchin early and late H2B histone gene expression by a gastrula nuclear extract after injection into Xenopus laevis oocytes.

Maxson¹,

Ito²,

Balcells³

et al. 1988

Mol. Cell. Biol.

View full text Add to dashboard Cite

Sea urchin early histone genes are active in preblastula embryos; late histone genes are maximafly expressed during subsequent stages of embryogenesis. We used the Xenopus laevis oocyte to assay for trans-acting factors involved in this differential regulation. Sea urchin nuclear proteins were prepared by extracting gastrula-stage chromatin successively with 0.45, 1, and 2 M NaCl. We injected three fractions into oocytes along with plasmids bearing sea urchin early and late H2b histone genes. While neither the 0 to 0.45 M nor the 1 to 2 M salt fraction affected H2b gene expression, the 0.45 to 1 M salt fraction stimulated early and late H2b mRNA levels significantly. Late H2b gene expression was stimulated preferentially when the early and late genes were coinjected into the same oocytes. This extract did not stimulate the accumulation of transcripts of injected herpesvirus thymidine kinase genes or of the sea urchin Spec 1 gene, suggesting that the stimulatory activity is not a general transcription factor. We localized the DNA sequence required for the stimulatory effect to a region of the late H2b gene located between -43 and +62 relative to the transcription start site. A component of the 0.45 to 1 M salt wash fraction specifically bound to the 105-base-pair late gene DNA sequence and to the corresponding early gene fragment. The abundance of this binding activity decreased on a per genome basis during early development of the sea urchin.The sea urchin synthesizes several distinct sets of histone proteins during early embryogenesis (7, 32). These include early isotypes, synthesized from fertilization to the blastula stage, and late isotypes, synthesized from the blastula stage onward. Each set of proteins is encoded by a distinct class of genes. Early histone genes are repeated several hundredfold in the genome and are organized in tandem quintets, each comprising genes for the five major histone types. In contrast, late histone genes are present in only 6 to 12 copies per genome and are arranged in irregular clusters (6,21,22,27). The developmental switches in the synthesis of early and late histone isotypes are caused largely by changes in rates of transcription of the two gene sets (23,28,35).The Xenopus laevis oocyte has proven highly useful in the analysis of transcriptional processes (9, 10). A variety of genes, including histone genes, are accurately transcribed after microinjection into oocyte nuclei, and the injection of genes with mutated promoter regions has enabled the identification of elements required for transcription in the oocyte (16). The Xenopus oocyte system has also been used to identify a trans-acting factor involved in the processing of sea urchin histone mRNAs (34) and, more recently, to identify a factor capable of stimulating sea urchin early H2b gene transcription (30).We used the Xenopus oocyte to examine the mechanisms underlying the differential expression of sea urchin early and late histone genes. We found that an extract of sea urchin gastrula-stage chromatin stimulated the ...

show abstract

“…The discovery of viral enhancer sequences (for a brief review, see reference 18) demonstrated that regulatory elements may also function outside of the more traditionally defined promoter region. It was subsequently demonstrated that a number of cellular genes (including those encoding immunoglobulin heavy and light chains [2,3,11,[28][29][30], human growth hormone [37], and sea urchin H2B [26]) possess internal regulatory elements. Several years ago we set out to investigate the existence of control elements internal to or downstream of goat p-globin genes.…”

mentioning

confidence: 99%

Differential expression of mouse beta/goat beta c, mouse beta/goat beta F, and mouse beta/goat epsilon II hybrid globin genes in murine erythroleukemia cells.

Kerlakian¹,

Tóth²,

Kuempel³

et al. 1986

Mol. Cell. Biol.

View full text Add to dashboard Cite

We assembled three hybrid 13-globin (10,12,13,52), and the heat shock-responsive element of the Drosophila hsp-70 gene (9,27 MATERIALS AND METHODSConstruction of fusion genes. We assembled three p-globin fusion genes for this study (Fig. 1) 'n20-kb' 1 jig of HSV-106, enough globin plasmid to give a 10-fold molar excess over HSV-106, and the remainder as highmolecular-weight purified F4-12B2 cell DNA. In the experiments with pUCtk-r, the plasmid was cleaved with ScaI g3C before transformation (Fig. 2); either 1.8 or 9 ,ug of the * pUCtk-e plasmid per plate was used. As in the earlier studies, F4-12B2 DNA was added so that a total of 25 ,ug of DNA was applied to each plate. Regardless of which plasmids were used for transformation, the DNA precipitate was left on the cells for 9 h, when they were fed with nonselective D medium. Twenty hours later, the initial medium was replaced with selective medium (HAT; 15 ,ug of hypoxanthine per ml, 0.2 jiM aminopterin, and 5 jig of thymidine per ml). . DNAs from other lines were dot blotted. Dot blots were or P3F). The mouse prepared by suspending the DNAs in 5 ,ul of 0.1 M NaOH-2 ,B/human 1 construct was that described by Chao et al. (6) and was kindly provided by that laboratory.After our initial round of experiments with the three constructs shown in Fig. 1, we wanted to test the effect of placing the mouse 13/goat Ell gene on the same DNA fragment as the selectable marker gene for thymidine kinase (tk). To accomplish this we first prepared the 1.8-kilobase (kb) PvuII fragment from herpes simplex virus 106 (HSV-106) (24) which contains the entire herpesvirus tk gene; this was cloned by blunt-end ligation into the unique SspI site of pUC18. The EcoRI fragment containing the mouse 1/goat Ell gene was then cloned into the BamHI site in the polylinker of the pUCtk plasmid by blunt-end ligation, creating plasmid pUCtk-r. This construct is diagrammed in Fig. 2.Introduction of globin genes into MEL cells. The F4-12B2

show abstract

Stimulation of sea urchin H2B histone gene transcription by a chromatin-associated protein fraction depends on gene sequences downstream of the transcription start site.

Cited by 35 publications

References 27 publications

Different Potential of Cellular and Viral Activators of Transcription Revealed in Oocytes and Early Embryos ofXenopus laevis

Different Potential of Cellular and Viral Activators of Transcription Revealed in Oocytes and Early Embryos ofXenopus laevis

Differential stimulation of sea urchin early and late H2B histone gene expression by a gastrula nuclear extract after injection into Xenopus laevis oocytes.

Differential expression of mouse beta/goat beta c, mouse beta/goat beta F, and mouse beta/goat epsilon II hybrid globin genes in murine erythroleukemia cells.

Contact Info

Product

Resources

About