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P Fransz

Publications and source records attributed to P Fransz.

11 recordsLinked to original sources
PubMed2006

Composition and formation of heterochromatin in Arabidopsis thaliana.

The term heterochromatin has been applied to both large-scale, microscopically visible chromocentres and small-scale, silent genes located outside chromocentres. This may cause confusion in the interpretation of epigenetic marks for both features. The model plant Arabidopsis thaliana provides an excellent system to investigate composition and function of chromatin states at different levels of organization. In this review we will discuss recent developments in molecular networks underlying gene silencing and the relationship with visible heterochromatin in Arabidopsis.

Arabidopsis↗
PubMed2003

Heterochromatin in interphase nuclei of Arabidopsis thaliana.

The eukaryotic nucleus represents a complex arrangement of heterochromatic and euchromatic domains, each with their specific nuclear functions. Somatic cells of a multicellular organism are genetically identical, yet they may differ completely in nuclear organization and gene expression patterns. Stable changes in gene expression without modifying the sequence are the result of epigenetic changes and include covalent modifications in cytosine residues of DNA and in histone tails giving rise to altered chromatin protein complexes, remodeling of chromatin and changes in chromatin compaction. Large-scale differences in chromatin structure are visible at the microscopic level as euchromatin and heterochromatin. Arabidopsis thaliana chromosomes display a relatively simple distribution of euchromatic and heterochromatic segments overlapping with gene-rich and repeat-rich regions, respectively. Recently, we have shown that Arabidopsis provides a well-defined system to study individual chromosomes and chromatin domains in interphase nuclei as well as the relationship between chromatin condensation and epigenetic mechanisms of gene silencing. This overview focuses on the organization and composition of heterochromatin in Arabidopsis nuclei.

Arabidopsis↗
PubMed2001

The centromere1 (CEN1) region of Arabidopsis thaliana: architecture and functional impact of chromatin.

We have analysed the centromere 1 (CEN1) of Arabidopsis thaliana by integration of genetic, sequence and fluorescence in situ hybridisation (FISH) data. CEN1 is considered to include the centromeric core and the flanking left and right pericentromeric regions, which are distinct parts by structural and/or functional properties. CEN1 pericentromeres are composed of different dispersed repetitive elements, sometimes interrupted by functional genes. In contrast the CEN1 core is more uniformly structured harbouring only two different repeats. The presented analysis reveals aspects concerning distribution and effects of the uniformly shaped heterochromatin, which covers all CEN1 regions. A lethal mutation tightly linked to CEN1 enabled us to measure recombination frequencies within the heterochromatin in detail. In the left pericentromere, the change from eu- to heterochromatin is accompanied by a gradual change in sequence composition but by an extreme change in recombination frequency (from normal to 53-fold decrease) which takes place within a small region spanning 15 kb. Generally, heterochromatin is known to suppress recombination. However, the same analysis reveals that left and right pericentromere, though similar in sequence composition, differ markedly in suppression (53-fold versus 10-fold). The centromeric core exhibits at least 200-fold if not complete suppression. We discuss whether differences in (fine) composition reflect quantitative and qualitative differences in binding sites for heterochromatin proteins and in turn render different functional properties. Based on the presented data we estimate the sizes of Arabidopsis centromeres. These are typical for regional centromeres of higher eukaryotes and range from 4.4 Mb (CEN1) to 3.55 Mb (CEN4).

Arabidopsis↗
PubMed2000

Endogenous targets of transcriptional gene silencing in Arabidopsis.

Transcriptional gene silencing (TGS) frequently inactivates foreign genes integrated into plant genomes but very likely also suppresses an unknown subset of chromosomal information. Accordingly, RNA analysis of mutants impaired in silencing should uncover endogenous targets of this epigenetic regulation. We compared transcripts from wild-type Arabidopsis carrying a silent transgene with RNA from an isogenic transgene-expressing TGS mutant. Two cDNA clones were identified representing endogenous RNA expressed only in the mutant. The synthesis of these RNAs was found to be released in several mutants affected in TGS, implying that TGS in general and not a particular mutation controls the transcriptional activity of their templates. Detailed analysis revealed that the two clones are part of longer transcripts termed TSI (for transcriptionally silent information). Two major classes of related TSI transcripts were found in a mutant cDNA library. They are synthesized from repeats present in heterochromatic pericentromeric regions of Arabidopsis chromosomes. These repeats share sequence homology with the 3' terminal part of the putative retrotransposon Athila. However, the transcriptional activation does not include the transposon itself and does not promote its movement. There is no evidence for a general release of silencing from retroelements. Thus, foreign genes in plants encounter the epigenetic control normally directed, at least in part, toward a subset of pericentromeric repeats.

Arabidopsis↗
PubMed1999

High resolution FISH in plants - techniques and applications.

Fluorescence in situ hybridization (FISH) is an effective and accurate cytogenetic tool for mapping single copy and repetitive DNA sequences on chromosomes. Attempts to increase the detection sensitivity of very small chromosomal targets, and to improve the spatial resolution of signals derived from flanking sequences, have led to the development of a variety of novel techniques: it is now possible to perform in situ hybridizations on interphase nuclei, meiotic pachytene chromosomes and isolated chromatin (DNA fibres). The recent application of these techniques has indicated that a spatial resolution of 1 kb between adjacent targets and a sensitivity of targets smaller than 1 kb is now feasible. Here, we describe the benefits of these novel chromosome analysis techniques and discuss their relevance for the study of plant genomes.

Journal Article↗
PubMed1999

The DIF1 gene of Arabidopsis is required for meiotic chromosome segregation and belongs to the REC8/RAD21 cohesin gene family.

Cohesins are a group of conserved proteins responsible for cohesion between replicated sister chromatids during mitosis and meiosis and which are implicated in double-strand break repair and meiotic recombination. We describe here the identification and characterisation of an Arabidopsis gene - DETERMINATE, INFERTILE1 (DIF1), which is a homolog of the Schizosaccharomyces pombe REC8/RAD21 cohesin genes, and is essential for meiotic chromosome segregation. Five independent alleles of the DIF1 gene were isolated by transposon mutagenesis, and the mutants show complete male and female sterility. Pollen mother cells (PMCs) of dif1 mutants show multiple meiotic defects which are represented by univalent chromosomes and chromosome fragmentation at metaphase I, and acentric fragments and chromatin bridges in meiosis I and II. Consequently, chromosome segregation is strongly affected, resulting in meiotic products of uneven size, shape and of variable ploidy. The similarities in phenotype, and the sequence homology between DIF1 and the REC8/RAD21 cohesins suggests that cohesin function is largely conserved between eukaryotes and highlights the essential role cohesins play in plant meiosis.

Alleles↗
PubMed1998

The behaviour of the autonomous maize transposable element En/Spm in Arabidopsis thaliana allows efficient mutagenesis.

The behavior of the autonomous maize transposable element En/Spm of maize was studied in Arabidopsis. Transgenic Arabidopsis plants carrying En-1 elements were propagated for 12 generations using a single seed descent procedure. The distribution and activity of the En-1 element was monitored using Southern DNA hybridisations in generations 1, 6 and 12. In the first generation the highest number of En-1 insertions per line was 7, which increased to 20 in generation 12. The average number of En-1 insertions increased only slightly in the population, due to a gradual accumulation of segregants that lost the transposable element. During the development of the En-1 mutagenised population the element remained active even in the high-copy lines. In situ hybridisation demonstrated that multiple En-1 insertions were distributed over all Arabidopsis chromosomes. From the initial En-1 mutagenised populations many unstable gene mutations were recovered, indicating that En-1 can be used as a efficient tool for gene tagging in Arabidopsis.

Arabidopsis↗
PubMed1998

Cytogenetics for the model system Arabidopsis thaliana.

A detailed karyotype of Arabidopsis thaliana is presented using meiotic pachytene cells in combination with fluorescence in situ hybridization. The lengths of the five pachytene bivalents varied between 50 and 80 microns, which is 20-25 times longer than mitotic metaphase chromosomes. The analysis confirms that the two longest chromosomes (1 and 5) are metacentric and the two shortest chromosomes (2 and 4) are acrocentric and carry NORs subterminally in their short arms, while chromosome 3 is submetacentric and medium sized. Detailed mapping of the centromere position further revealed that the length variation between the pachytene bivalents comes from the short arms. Individual chromosomes were unambiguously identified by their combinations of relative lengths, arm-ratios, presence of NOR knobs and FISH signals with a 5S rDNA probe and chromosome specific DNA probes. Polymorphisms were found among six ecotypes with respect to the number and map positions of 5S rDNA loci. All ecotypes contain 5S rDNA in the short arms of chromosomes 4 and 5. Three different patterns were observed regarding the presence and position of a 5S rDNA locus on chromosome 3. Repetitive DNA clones enabled us to subdivide the pericentromeric heterochromatin into a central domain, characterized by pAL1 and 106B repeats, which accommodate the functional centromere and two flanking domains, characterized by the 17 A20 repeat sequences. The upper flanking domains of chromosomes 4 and 5, and in some ecotypes also chromosome 3, contain a 5S rDNA locus. The detection of unique cosmids and YAC sequences demonstrates that detailed physical mapping of Arabidopsis chromosomes by cytogenetic techniques is feasible. Together with the presented karyotype this makes Arabidopsis a model system for detailed cytogenetic mapping.

Arabidopsis↗
PubMed1997

Cytological characterization of four meiotic mutants of Arabidopsis isolated from T-DNA-transformed lines.

A secondary screen of the Feldmann collection of T-DNA transformed Arabidopsis lines identified several meiotic mutants. We used a spreading technique combined with DAPI staining in a detailed cytogenetic analysis of meiotic chromosome behaviour in four of these mutants, all of which are putatively T-DNA tagged and therefore candidates for molecular and functional analysis of the mutated genes. Two of them are defined as 'synaptic' mutants, showing greatly reduced association of homologous chromosomes at metaphase I: one is asynaptic, showing failure of synapsis during prophase I, whereas the other is desynaptic and is characterized by normal but non-maintained synapsis. Another mutant is defective in meiotic cell cycle control and undergoes a third meiotic division, resembling a second division but without an additional round of chromosome duplication. A further mutant shows meiosis-limited chromosome disruption, resulting in extensive chromosome fragmentation combined with other defects. All four mutants experience very irregular chromosome distribution during the meiotic divisions, resulting in abnormal numbers and/or sizes of microspores, with resulting reduced fertility.

Arabidopsis↗
PubMed1996

A light microscopic atlas of meiosis in Arabidopsis thaliana.

An atlas of meiosis in Arabidopsis thaliana, encompassing all stages from preleptotene to telophase II and early microspore formation, is presented in detail for the first time. The photomicrographs and descriptions are based on staining with the DNA fluorochrome 4',6-diamidino-2-phenylindole (DAPI) combined with a spreading procedure, or haematoxylin-iron alum (HIA) staining. Despite previous reservations about the practicality of cytogenetic meiotic analysis in Arabidopsis due to its small genome size, good-quality and clearly analysable preparations of all meiotic stages were obtained. This atlas of normal, wild-type meiosis is considered an essential prerequisite to informed analyses of meiotic mutants. Furthermore, extended prophase I chromosomes, particularly at the pachytene stage, offer considerable potential for producing a detailed cytogenetic map (karyotype) of Arabidopsis chromosomes with the additional prospect of high-resolution physical mapping based on fluorescence in situ hybridization (FISH) of defined DNA probes to these extended chromosomes.

Anaphase↗