Spelling suggestions: "subject:"parameterization"" "subject:"parameterisation""
1 |
Tandem Repeats are Sufficient for b1 ParamutationBelele, Christiane January 2006 (has links)
Paramutation is an allele interaction that causes a heritable change in the expression of one allele. At the b1 locus an interaction between B' and B-I alleles results in a change of B-I to B', symbolized by B'*. A combination of fine-structure mapping and transgenic approaches have demonstrated that the tandem repeats located ~100 kb upstream of the b1 transcription start site are sufficient for both paramutation and high expression.Plants carrying transgenes with tandem repeats in ectopic locations (repeat-transgene) were able to change B-I into B'*. The B'* state induced by the repeat-transgene was heritable and paramutagenic when segregated from the repeat-transgene. In addition, the repeat-transgene induced B-I silencing was prevented by the trans-acting mutation required for paramutation mop1-1, which was recently found to encode a RNA-dependent RNA polymerase (RdRP). Transgenes containing seven tandem repeats of only the 5' half of the sequence were able to paramutate B-I. Taken together, these results demonstrate that the paramutation sequences are contained in the 5' half of the repeats and they can paramutate B-I from non-allelic positions. Because paramutation induced by the repeat-transgenes and the endogenous B' allele are both heritable and depend on a functional RdRP, they likely involve a similar mechanism of RNA-mediated chromatin modification.Furthermore, we found that the tandem repeats are also sufficient for high expression of the b1 gene. When fused to a GUS reporter gene and introduced into maize, the tandem repeats enhanced GUS expression above the level observed for GUS transgenes that did not have the repeats. As observed with the endogenous B-I allele, the enhancer function of the repeats in the GUS transgenes is silenced by B' and the paramutagenic repeat-transgenes. After being with B' or the paramutagenic repeat-transgenes the repeats in the GUS constructs lost their ability to enhance gene expression.The identification of the tandem repeats as the sequences mediating paramutation suggest a new function for tandem repeats, mediating trans-interactions to establish heritable epigenetic states. Models are discussed for how alleles might communicate in trans to establish different epigenetic states and how the epigenetic state is maintained through mitosis and meiosis.
|
2 |
Emergence d'un locus producteur de piRNAs chez la drosophile : mise en place de l'épigénome / Emergence of a piRNA-producing locus in drosophilaHermant, Catherine 28 January 2015 (has links)
Les éléments transposables d’ADN sont presque ubiquitaires dans le monde vivant et leur mobilité peut être délétère pour le génome. Leur régulation dans les tissus germinaux animaux passe par la voie de silencing des piRNAs (PIWI-interacting RNAs). Les piRNAs sont produits à partir de loci contenant des fragments d’éléments transposables insérés en clusters. Nous étudions l’émergence de ces clusters de piRNAs chez la drosophile.Nous avons activé de novo un cluster de transgènes par héritage maternel de piRNAs homologues. Il s’agit d’un cas de paramutation, ou conversion épigénétique stable et récurrente.Nous avons montré que ce cluster paramuté produit de novo des piRNAs, et étonnement dessiRNAs.J’ai caractérisé de façon fonctionnelle et moléculaire ce phénomène de paramutation par l’utilisation de mutants. J’ai montré que les propriétés de silencing, ainsi que la production depiRNAs et de siRNAs, sont abolies en contexte mutant pour tous les gènes testés de la voie despiRNAs (voies primaire et secondaire). Parallèlement, j’ai étudié un cas de paramutation« partiellement homologue » dans laquelle le cluster reçoit des piRNAs homologues seulement àune partie de sa séquence. J’ai montré qu’il y a production de piRNAs par la totalité du cluster dès la 3e génération.J’ai montré, enfin, que des clusters activés de novo par la chaleur, présentent des propriétés fonctionnelles et moléculaires semblables aux clusters activés par les piRNAsmaternels.Ces travaux apportent des éléments clés pour la compréhension de la mise en place de l’épigénome, tant d’un point de vue mécanistique qu’évolutif. / DNA transposable elements are almost ubiquitous in the living world and their mobility can be deleterious for the genome. Their regulation in germaria is mediated by the piRNAsilencing pathway (PIWI-interacting RNAs). piRNAs are produced by loci formed by clusters of fragments of transposable elements. We are studying the emergence of these piRNA-producing clusters in Drosophila.We have de novo activated a cluster of transgenes via maternal inheritance of homologous piRNAs. This is a case of paramutation i.e. a stable and recurrent epigenetic conversion process.We have shown that this paramutated cluster produces de novo piRNAs and, surprisingly, also siRNAs. I have characterized this paramutation functionally and molecularly, by a mutant approach. I have shown that its silencing properties, as well as piRNA and siRNA production are abolished in mutant contexts for all the genes from the primary and secondary piRNA pathways I have tested. At the same time, I have studied the case of a partially homologous paramutation, in which piRNAs maternally inherited by the cluster are homologous to only a part of its sequence. I have shown that piRNA are produced all along the cluster as early as the 3rdgeneration.Finally, I have shown that a cluster activated de novo by an environmental stress shows the same functional and molecular properties as a cluster paramutated via maternal piRNA inheritance.These studies provide key elements for understanding the emergence of the epigenome from a mechanistic and an evolutionary perspective.
|
3 |
Molecular Characterization of the mop2, a Gene Required for Epigenetic SilencingCai, Yu January 2006 (has links)
The mop2 gene is required for epigenetic silencing; it was originally defined as a mutation, Mop2-1, which when dominant prevented paramutation at b1. Paramutation is an allele communication that causes a mitotically and meiotically heritable change in gene expression. Mop2-1 was subsequently shown to be involved in maintaining the silenced paramutant state and to prevent dsRNA-mediated transcriptional gene silencing (activities revealed only when the mutation is homozygous). Understanding the product encoded by mop2 will help dissect the underlying mechanisms involved in paramutation and dsRNA-mediated transcriptional silencing. This dissertation describes map-based cloning and candidate gene approaches directed toward the eventual goal of identification of mop2.Initial mapping of mop2 placed it within a region delineated by the markers umc1823 and eks1. On the maize physical map this region contains 21 BAC (Bacteria Artificial Chromosome) clones, representing 2.9 Mb. Skim sequencing identified additional markers for mapping and revealed the gene content. Extensive candidate gene examinations, including gene sequencing, expression profiling with microarrays and RT-PCR, and complementation tests with mutant alleles did not identify any of the four chromatin and RNAi-related genes as mop2.The new markers developed from the skim sequence enabled further mapping and molecular genotyping, which revealed that the Mop2-1 mutation was unstable. Approxi¬mately 10% of phenotypic heterozygous plants were actually genotypic homozygous. Further mapping using only Mop2-1 homozygous plants reduced the mop2 interval to a region of nine BACs, containing 57 genes.The mop2 region is highly syntenic to a rice region of 1.25 Mb on chromosome 4. The gene alignment and repetitive sequence analyses between the syntenic regions in these two species revealed both syntenic and non-syntenic blocks of sequences. Analyses suggested several potential mechanisms for the collinearity breakage, including, but not limited to, tandem duplications of genes in one species but not the other and the presence of gene fragments in maize, but not in rice.The research described herein provides the basis for continued efforts to clone mop2. Fine-structure mapping with new markers and a larger population, as well as candidate gene sequencing in the Mop2-1 BAC library, should be pursued to clone mop2.
|
4 |
INTER-KINGDOM EPIGENETICS: CHARACTERIZATION OF MAIZE B1 TANDEM REPEAT-MEDIATED SILENCING IN DROSOPHILA MELANOGASTERMcEachern, Lori A. 19 August 2010 (has links)
Transgenic organisms are a valuable tool for studying epigenetics, as they provide significant insight into the evolutionary conservation of epigenetic control sequences, the interacting proteins, and the underlying molecular mechanisms. Paramutation is an epigenetic phenomenon in which the epigenetic status and expression level of one allele is heritably altered after pairing with another. At the b1 locus in maize, a control region consisting of seven 853 bp tandem repeats is required for paramutation. To study the conservation of the epigenetic mechanisms underlying maize b1 paramutation, I created transgenic Drosophila carrying the maize b1 control region flanked by FRT sites and adjacent to the Drosophila white reporter gene. The maize b1 tandem repeats caused epigenetic silencing in Drosophila, as white expression consistently increased following repeat removal. A single copy of the tandem repeat sequence was sufficient to cause silencing, and silencing strength increased as the number of repeats increased. Trans interactions, such as pairing-sensitive silencing, were also observed and appear to require a threshold number of b1 tandem repeats, similar to paramutation in maize. Analysis of transcription from the repeats showed that the b1 tandem repeats are transcribed from both strands in Drosophila, as they are in maize. Bidirectional transcription was found to extend to the regions flanking the repeats, and persisted in “repeats-out” transgenes following repeat removal. However, aberrant transcription was lost when a zero-repeat transgene was moved to a new genomic position, suggesting that it may be due to an epigenetic mark that is retained from the previous silenced state. A search for modifiers of b1 repeat-mediated silencing demonstrated that Polycomb group proteins are involved. Together, these results indicate considerable conservation of an epigenetic silencing process between the plant and animal kingdoms. Genomic imprinting is a related epigenetic process in which parent-specific epigenetic states are inherited and maintained in progeny. The conservation of epigenetic mechanisms was further explored via an in-depth review of the molecular mechanisms underlying genomic imprinting in plants, mammals and insects, and identification of potentially imprinted genes in Drosophila by microarray analysis.
|
5 |
Molecular mechanisms that underlie non-Mendelian inheritance patterns in <i>Zea mays</i>Deans, Natalie Christine January 2020 (has links)
No description available.
|
Page generated in 0.0944 seconds