Jean-Pierre Jost

Emeritus / Friedrich Miescher Institute for Biomedical Research · gene regulation

Quatrième de couverture Saviez-vous que l’ours nous a appris à utiliser certaines plantes médicinales, que la marmotte a permis d’optimiser les transplantations ou encore que les champignons peuvent être bien utiles pour lutter contre la sécheresse et la pollution ? Depuis toujours, les sociétés humaines ont observé attentivement le comportement an...

Summary Plants communicate with each other in many ways. They signal their presence to their neighbours by emitting infrared light. This allows the latter to avoid their shadow. They are not deaf, they perceive and emit sound waves. In response to certain frequencies, they selectively activate several genes (resistance to insects and diseases, phot...

Les insectes sociaux tels que fourmis, termites, abeilles, guêpes, et frelons maitrisent soit l’usage de l’argile, soit de la cire ou du cartonnage pour la construction de leurs demeures. Le système d’air conditionné des termites inspira les architectes modernes de même que les structures hexagonales des rayons d’abeilles furent adoptées par les in...

Quatrième de couverture Les animaux sont en mesure de nous donner de bons exemples d’entraide, d’altruisme et parfois même d’empathie. Dans la nature l’entraide est omniprésente et c’est l’un des moteurs important de l’évolution des espèces. L’entraide s’observe aussi bien chez les insectes sociaux que chez les mammifères supérieurs et certains d’e...

Quatrième de couverture Les animaux sont en mesure de nous donner de bons exemples d’entraide, d’altruisme et parfois même d’empathie. Dans la nature l’entraide est omniprésente et c’est l’un des moteurs important de l’évolution des espèces. L’entraide s’observe aussi bien chez les insectes sociaux que chez les mammifères supérieurs et certains d’e...

Fourth cover This book presents several ecological processes capable of limiting the use of insecticides that pollute the soil and water. Among the different strategies there are: cultural methods such as crop rotation, catch and intercropping and the date of sowing which contribute to the fight against harmful insects and nematodes. The classic se...

The original text is in French. Summary: Communication among social and non-social insects is done through four basic types of signals: sound, tactile, visual and chemical (pheromones). Due to the complexity of the interactions in social insects they appeal to several different communication channels. This is the case in bees, ants, termites and co...

Summary Plants hyperaccumulating heavy metals such as Zn, Cu, Pb, Cr, Se, As, Hg, Cd, Ni, Ag, Au, or Pt are capable of absorbing large quantities of them; they turn out to be useful for decontaminating polluted soil and water. To date we know approximately 1000 different plants capable of tolerating or hyper accumulating heavy metals and each plant...

This book is an essay on how plants communicate to each other to signal their presence.They do it by the quality of light (infrared) emitted by their leaves. They are capable to warn their neighbors about impeding dangers (diseases, predator). This is achieved by means of stress hormones (jasmonate, salicylate and ethylene) and other volatile chemi...

For the past 500 million years transgenesis was also instrumental to the evolution of species. In the past 40 years scientists made use of transgenesis to create or improve many cultivars. Recently the edition of genome enables to target with high precision new genes into the genome of plants. This technique permits also to replace and repair genes...

Plants have many ways to prevent or to respond to attacks by insects, viruses, bacteria or fungae. They are able to modify the structures of the cell walls or modify the number of stomata and thorns. They respond to elicitors and stress hormones by synthesizing special chemicals (phytoalexins) that are acting against insects and pathogens. They are...

5-methylcytosine DNA glycosylase (5-MCDG) and G/T mismatch DNA glycosylase activities are present in the same human recombinant protein. When tested with 2x 10 -8 M substrates (54% G+C) in the presence of the intercalating antibiotic Echinomycin, G/T mismatch DNA glycosylase is strongly inhibited with an apparent Ki of o.6 micro M while 5-MCDG acti...

From insects to elephants we observe self-medication. Chemical and physical properties of plants or soil are exploited to fight against various endo and exo parasites. Drosophila flies as well as elephants may abuse of alcohol in stress situations. Some animals become drug addicted. Steroid like substances produced in plants serve as contraceptives...

This book gives a summary about all what we know on the mechanisms of hibernation and estivation in general and it gives several examples among different animal species . It gives also a brief presentation on the possible applications in medicine. For further details look at www.jostwildlife.com (under books , summary), feb. 2014

Monograph on the European Wild Cat. Biology, Behavior, genetic analysis, territoriality, cross-breeding with domestic cats, reintroduction, distribution in Europe and protection

This book deals with: Communication between animals or animals and humans. Social learning, tool use, metacognition, notion of time, numbers and serial order. Long term and short term memory, spatial memory, associative learning, self recognition. Planing of future actions. Deceptive manoeuvres. Empathy toward humans. Over 60 explicative drawings a...

Short stories (36) about a boy ( the author) who was kicked out of school to work as a farm worker and postman in the Swiss alps and some other stories about this time (1950)

Monograph on the european Lynx. Zoology, behaviour, distribution in Europe, territoriality, reintroductions , protection, ecological role of the lynx, problems of cohabitation with humans.

Monograph on the European Beaver. Biology, behaviour, social structure, reintroduction, impact on ecosystems, management and protection

Instinct or intelligence? A review on intelligent behaviour of birds and mammals, cognitive capacity, tool use, altruism, empathy, communication. Collective intelligence by insects, robotic.

Some 106 different kind of birds come from the north or the east to spend the winter or part of it in Switzerland and surrounding countries. Its only a small part of the 5 billion birds that migrate every year to the south . The book describe where they come from and where spend the winter in Switzerland and when. Over 106 original drawings from th...

This monograph on the Red Deer deals with the origin, biology, social structure, hunting, importance in mythology and traditional medicine,Sort description of other members of the same family living in Europe. For further information: www.jostwildlife.com

Monograph on the return of the European brown Bear. Biology, behaviour,, reintroduction in different countries. Problems linked with its protection, scientific research, popular medicine and shamanism, is the bear a dangerous predator? the hibernation of the bear. A short summary on the different forms of hibernation and the estivation.

Monograph on the Ibex (Capra ibex) living in the Alps. The story of its reintroduction in Switzerland at the beginning of the 20th century. Biology, behaviour, social structure, reproduction and dispersion. Description of the other species living in the world. Details in www.jostwildlife.com

monograph on the chamois (Rupicapra rupicapra). Geographic distribution in Europe. Reintroductions and introductions in other countries. Biology, behaviour, hunting of.. The other subspecies of Rupicapra For further information see www.jostwildlife.com

Monograph on the Red Fox. Urban and peri-urban life of..., Biology, behavior, control of rabies, for further information look at www.jostwildlife.com

Monograph on the Alpine Marmots. Biology, behavior, territoriality, details on the hibernation. Twenty different species of marmots in the world. For further details look at www.jostwildlife.com

All about the social structure of the alpine marmots, territoriality, interactions with other animals, reproduction , the den (summer ans winter). Preparation for hibernation. Hibernation. Physiology of hibernation . Different kind of marmots around the world. Hunting , The marmots as an economical resource

At a concentration of 5 x 10(-9) M of hemi-methylated DNA (one order of magnitude below the K(m)), MCF-7 (a human breast carcinoma cell line) nuclear extracts potentiate the activity of 5-methylcytosine DNA glycosylase (5-MCDG, alias G/T mismatch DNA glycosylase). Depending on the ratio between MCF-7 nuclear extracts and 5-MCDG, there is an up to 1...

Human mature sperm cells have a high nuclease and 5-methyldeoxycytidine monophosphate (5-mdCMP) deaminase activity. The deaminase converts the nuclease degradation product 5-mdCMP into dTMP which is further cleaved into thymine and the abasic sugar-phosphate. Both 5-methylcytidine 5' and 3' monophosphates are good substrates for the deaminase. 5-me...

Changes in gene expression during mouse myoblast differentiation were monitored by DNA microarray hybridisation. Four days after the onset of differentiation 2.37% of the genes increased in activity from a value of zero, whereas during the same time 1.68% of total genes had decreased expression. During the first 24 h of differentiation an average o...

We have constructed a stable cell line, human embryonal kidney 293M+, containing a lacZ reporter gene controlled by an in vitro methylated hormone-responsive enhancer. Methylation of the enhancer–promoter abolishes lacZ expression controlled by ponasterone A (an analogue of ecdysone). Ponasterone A-induced expression is restored by the short-chain...

We have shown that the DNA demethylation complex isolated from chicken embryos has a G(.)T mismatch DNA glycosylase that also possesses 5-methylcytosine DNA glycosylase (5-MCDG) activity. Herein we show that human embryonic kidney cells stably transfected with 5-MCDG cDNA linked to a cytomegalovirus promoter overexpress 5-MCDG. A 15- to 20-fold ove...

A 1468 bp cDNA coding for the chicken homolog of the human MBD4 G/T mismatch DNA glycosylase was isolated and sequenced. The derived amino acid sequence (416 amino acids) shows 46% identity with the human MBD4 and the conserved catalytic region at the C-terminal end (170 amino acids) has 90% identity. The non-conserved region of the avian protein h...

In the developing chicken embryo, active DNA demethylation requires both RNA and proteins (Nucleic Acids Res. 25, 2375-2380, 1997; ibid. 25, 4545-4550, 1997, FEBS Lett. 449, 251-254, 1999a). In vitro assays indicate that in the 5- and 12-day-old embryos the highest specific activity of 5-methylcytosine DNA glycosylase is found in the brain, the eye...

We previously have shown that DNA demethylation by chicken embryo 5-methylcytosine DNA glycosylase (5-MCDG) needs both RNA and proteins. One of these proteins is a RNA helicase. Further peptides were sequenced, and three of them are identical to the mammalian G/T mismatch DNA glycosylase. A 3,233-bp cDNA coding for the chicken homologue of human G/...

The study of changes in genome-wide levels of DNA methylation has become a key focus for understanding the epigenetic regulation of gene expression. Many procedures exist to study DNA methylation, falling into two categories: gene-specific and genome-wide. Genome-wide methylation analysis is best performed by DNA hydrolysis followed by HPLC; howeve...

We have shown previously that DNA demethylation by chick embryo 5-methylcytosine (5-MeC)-DNA glycosylase needs both protein and RNA. Amino acid sequences of nine peptides derived from a highly purified 5-MeC-DNA glycosylase complex were identified by Nanoelectrospray ionisation mass spectrometry to be identical to the mammalian nuclear DEAD box pro...

Recently published results (Nucleic Acids Res. 26, 5573-5580, 1998) suggest that the ribonuclease sensitivity of the DNA demethylation reaction may be an experimental artifact due to the possible tight binding of the nucleases to the methylated DNA substrate. Using an improved protocol we show for two different systems that demethylation of hemimet...

Passive demethylation of DNA occurs through nucleotides analogs, carcinogens, antisense sequences or by drugs influencing the level of S-adenosylmethionine. Active DNA demethylation, whereby 5-methylcytosine is replaced by cytosine occurs by the action of 5 methylcytosine DNA glycosylase. Active DNA demethylation is also influenced by cic and trans...

A procedure for the efficient transfer of cell monolayers, cultured on glass coverslips, to microscopy slides has been developed. This technique involves the coating of the upper surface of an ethanol-fixed cultured cell layer with a film of collodion dissolved in n-amyl acetate. The dry collodion-coated cell layer can then be detached by rehydrati...

It has been shown that, during the S-phase of the cell cycle, the mouse DNA methyltransferase (DNA MTase) is targeted to sites of DNA replication by an amino acid sequence (aa 207-455) lying in the N-terminal domain of the enzyme [Leonhardt, H., Page, A. W., Weier, H. U. and Bestor, T. H. (1992) Cell , 71, 865-873]. In this paper it is shown, by us...

A 52 kDa protein (p52) was purified from chicken embryos and its corresponding cDNA was cloned. The p52 cDNA is 1768 bp long and has an open reading frame of 465 amino acids. The sequence of the p52 cDNA shows significant homology with mouse and human cDNAs from the EST database, so do the deduced amino acid sequences, indicating the existence of h...

The methylated DNA binding protein-2-H1 (MDBP-2-H1), present in rooster liver, is a member of the histone H1 family which inhibits transcription by binding selectively to methylated promoters. Here we have determined the primary structure of MDBP-2-H1. A comparison between histone H1 and MDBP-2-H1 was achieved by analyzing reversed phase HPLC-purif...

We have previously shown that DNA demethylation by chick embryo 5-methylcytosine (5-MeC)-DNA glycosylase needs both protein and RNA. RNA from enzyme purified by SDS-PAGE was isolated and cloned. The clones have an insert ranging from 240 to 670 bp and contained on average one CpG per 14 bases. All six clones tested had different sequences and did n...

Changes in DNA methylation during tobacco pollen development have been studied by confocal fluorescence microscopy using a monoclonal anti-5-methylcytosine (anti-m5C) antibody and a polyclonal anti-histone H1 (anti-histone) antibody as an internal standard. The specificity of the anti-m5C antibody was demonstrated by a titration series against both...

We have previously purified and characterized a 5-methylcytosine (5-MeC)-DNA glycosylase from 12 day old chick embryos [Jost,J.P. et al. (1995) J. Biol. Chem. 270, 9734–9739]. The activity of the purified enzyme is abolished upon treatment with proteinase K and ribonuclease A. RNA copurifies with 5-MeC-DNA glycosylase activity throughout all chroma...

Plant genomic DNA methylation was analyzed by an improved SssI methyltransferase assay and by genomic sequencing with sodium bisulfite. Kanamycin, hygromycin, and cefotaxime (also called Claforan) are commonly used as selective agents for the production of transgenic plants. These antibiotics caused DNA hypermethylation in tobacco plants grown in v...

We have detected sequence-specific non-symmetrical cytosine methylation within a 140 bp region of the promoter for the tobacco auxin-binding protein gene T85 in pollen DNA. Direct sequencing of the population of bisulphite reaction products showed that, in this region. 10 out of a possible 49 cytosine residues were methylated at a high frequency in...

Upon the onset of mouse myoblast differentiation there is a rapid drop in DNA methyltransferase activity followed by a genome wide demethylation [Jost and Jost (1994) J. Biol. Chem. 269, 10040–10043]. Here we show by using specific antibodies directed against DNA methyltransferase that upon differentiation there was a rapid drop in nuclear DNA meth...

We have previously shown that in vivo estradiol-dependent dephosphorylation of MDBP-2-H1 (a member of the histone H1 family) correlates with the loss of in vitro preferential binding to methylated DNA. To study the effects of the phosphorylation/dephosphorylation of MDBP-2-H1 on the expression of the avian vitellogenin II gene, we optimised an in v...

We have previously purified and characterized a ubiquitous non-histone protein (NHP1) which has a high affinity (Kd 10(-11) M) for different avian vitellogenin gene sequences containing CpGs (Hughes et al. (1989) Biochemistry 28, 9137-9142; Hughes and Jost (1989) Nucleic Acids Res. 17, 8511-8520). Here we show by microsequencing that the peptides d...

The expression of the serine protease inhibitor Protease nexin-1 (PN-1) is upregulated in glial cells following different types of lesion in the nervous system. A strong negative regulatory element has been shown by the missing nucleoside technique to be a CACGTG site (E-box) in the proximal part of the PN-1 promoter. The factor binding to this sit...

By using a purified fraction of mouse DNA methyltransferase we have shown, by gel-retardation analysis, that the enzyme forms a low-affinity complex preferentially with hemimethylated DNA; the complexes formed with unmethylated or with fully methylated DNA are of even lower affinity, and only very weak interaction occurs with DNA lacking CG dinucle...

Incubation of hemimethylated and labelled oligodeoxynucleotides with nuclear extracts from differentiating chicken embryos and mouse myoblasts resulted in the replacement of m5C by C. One of the enzymes involved is m5CpG endonuclease. It cleaves only m5CpG and not, m5CpT, m5CpA, m5CpC or m6ApT. The enzyme is not sequence specific and catalyses the...

PCR (polymerase chain reaction) is now one of the most widely used of basic molecular biology techniques and is an indispensable research tool for the molecular biologist. The basic PCR technique provides the cornerstone for in vitro DNA amplification - allowing the researcher to produce large quantities of DNA from minute amounts of starting mater...

We have previously shown that estradiol treatment of roosters resulted in a rapid loss of binding activity of the repressor MDBP-2-H1 (a member of the histone H1 family) to methylated DNA that was not due to a decrease in MDBP-2-H1 concentration. Here we demonstrate that MDBP-2-H1 from rooster liver nuclear extracts is a phosphoprotein. Phosphoamin...

We have previously shown that in developing chicken embryos and differentiating mouse myoblasts, the demethylation of 5-metCpGs occurs through the replacement of 5-methylcytosine by cytosine (Jost, J. P. (1993) Proc. Natl. Acad. Sci. U.S.A. 90, 4685-4688; Jost, J. P. & Jost, Y.C. (1994) J. Biol. Chem. 269, 10040-10043). We have now purified over 30...

The bisulfite method is a highly sensitive approach to 5-methylcytosine mapping that utilizes the capability of the polymerase chain reaction to exponentially amplify DNA. We have observed that the bisulfite reaction results in a significant level of template degradation due to DNA depurination. Furthermore, our data suggest that the DNA fragmentat...

DNA methyltransferase forms a low affinity complex preferentially with hemimethylated DNA. Only very weak interactions are observed with DNA lacking CpGs. Interaction is inhibited by N ethylmaleimide. Study on the sensitivity of the complex to proteolytic enzymes, Dynamic of reactions of methyl transfer etc.

This chapter discusses in vivo genomic footprinting with dimethyl sulfate (DMS). In vivo footprinting with DMS is a powerful tool to study the in situ interaction of proteins with DNA in intact cells. The major advantage of using DMS lies in the fact that because of its small molecular weight it readily penetrates cells, where it reacts with unprot...

We show that in mouse myoblasts the MyoD1 promoter is highly stimulated by MyoD1 expression, suggesting that it is controlled by a positive feedback loop. Using deletion and mutation analyses, we identified the targets for MyoD1 promoter autoregulation as the two proximal E-boxes located close to the MyoD1 core promoter. Gel mobility shift competit...

It has been recently shown that in developing chicken embryonic nuclear extracts there is a 5-methyldeoxycytidine excision repair activity (Jost, J. P. (1993) Proc. Natl. Acad. Sci. U. S. A. 90, 4684-4688). We show that in differentiating mouse myoblasts, a similar enzymatic reaction may be responsible for the genome-wide DNA demethylation (up to 5...

We have studied the transcriptional activity of the mouse MyoD1 gene promoter in vivo and in vitro using mouse G8 myoblasts and muscle cell nuclear extracts. 5' deletion analysis of the promoter and transcription-competition analysis using oligonucleotides corresponding to several cis-acting elements revealed that the basal activity of the MyoD1 pr...

Genomic footprinting is an extension of the genomic sequencing technique and is used to study protein/DNA interactions in vivo 1) (Fig. 1) It gives information concerning the sequence of protein binding elements and the contact points between a protein and its DNA target extend. Proteins bound to the DNA alter the reactivity of the bases towards UV...

Here I show that nuclear extracts of chicken embryos can promote the active demethylation of DNA. The evidence shows that in hemimethylated DNA (i.e., methylated on one strand only) demethylation of 5mCpG occurs through nucleotide excision repair. The first step of demethylation is the formation of specific nicks 5' from 5-methyldeoxycytidine. Nick...

With the availability of direct genomic footprinting techniques the study of native genomes has been greatly facilitated. This review provides an overview of the techniques involved and gives also a description of the mode of action of different DNA modifying agents which can be used for such methods. These include exonuclease III, deoxyribonucleas...

The major aim of this chapter is to give a brief overview of the most commonly used procedures to study minor DNA bases, especially 5-methylcytosine, and to refer the interested reader to the relevant methodological literature.

Steroid hormones are known to be directly involved in the differentiation of organs and tissues and to modulate the expression of specific genes. For the differentiation of specific tissues, methylation and demethylation of DNA may play an important role (Holliday, 1990), whereas for the modulation of gene expression specific steroid receptors rath...

MDBP-2 is a repressor that binds preferentially to methylated DNA. Peptides derived from MDBP-2 were sequenced. The sequences of the two peptides, KPAGPS-VTELITK and ALAAGGYDVEK, are identical to those found in the chicken histone H1 core protein. In SDS/polyacrylamide gels MDBP-2 has an apparent molecular mass of 21 kDa, and antibodies directed ag...

We have isolated the mouse MyoD1 gene flanked by its promoter region by screening a genomic library with synthetic ligonucleotides. The structural gene is interrupted by two G + C rich introns. Transfection of the cloned gene inserted into an expression vector converts fibroblasts to myoblasts. Sequence analysis of about 650 bp of the 5′ upstream r...

A negative regulating protein (MDBP-2) from rooster liver nuclear extracts binds preferentially to a methylated promoter region 5'TTCACCTTmCGCTATGAGGGGGATCATACTGG3' of the avian vitellogenin II gene (Nucleic Acids Res. 19, 1029-1034, 1991). Treatment of adult and immature roosters with estradiol results in a 90% decrease in the binding activity of...

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