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Dissertation
Functional Analysis of Src42A in embryonic epithelial morphogenesis of Drosophila melanogaster
(2023)
Src42A is one of the Src family kinases (SFKs) present in Drosophila melanogaster. In mammals there are 8 SFKs and so far, a loss of function analysis of SFKs were difficult to conduct due to their functional redundancy. In Drosophila, there are only two SFKs namely Src42A and Src64B, the former is essential for embryonic development and the latter is a non-essential gene. Since Src42A is supplied as a maternal component, the zygotic mutant were showing defects in later developmental stages, because the contribution ...
Aufsatz
A simple retroelement based knock-down system in Dictyostelium
(PLoS, 2015)
Characteristics of DIRS-1 Mediated Knock-Downs __ We have previously shown that the most abundant Dictyostelium discoideum retroelement DIRS-1 is suppressed by RNAi mechanisms. Here we provide evidence that both inverted terminal repeats have strong promoter activity and that bidirectional expression apparently generates a substrate for Dicer. A cassette containing the inverted terminal repeats and a fragment of a gene of interest was sufficient to activate the RNAi response, resulting in the generation of ~21 nt ...
Aufsatz
SSPIM: a beam shaping toolbox for structured selective plane illumination microscopy
(2018-07-03)
Selective plane illumination microscopy (SPIM) represents a preferred method in dynamic tissue imaging, because it combines high spatiotemporal resolution with low phototoxicity. The OpenSPIM system was developed to provide an accessible and flexible microscope set-up for non-specialist users. Here, we report Structured SPIM (SSPIM), which offers an open-source, user-friendly and compact toolbox for beam shaping to be applied within the OpenSPIM platform. SSPIM is able to generate digital patterns for a wide range ...
Aufsatz
Requirement of the Dynein-Adaptor Spindly for Mitotic and Post-Mitotic Functions in Drosophila
(2018-03-30)
Spindly was originally identified as a specific regulator of Dynein activity at the kinetochore. In early prometaphase, Spindly recruits the Dynein/Dynactin complex, promoting the establishment of stable kinetochore-microtubule interactions and progression into anaphase. While details of Spindly function in mitosis have been worked out in cultured human cells and in the C. elegans zygote, the function of Spindly within the context of an organism has not yet been addressed. Here, we present loss- and gain-of-function ...
Dissertation
Development of digital fluorescence light sheet microscopy for high-resolution optical imaging
(2021-12-15)
In modern biology investigating the morphological development of an organism is an inevitable task. Optical microscopy has been extensively used to monitor the live tissue changes in a model organism and it is typically used in conjunction with white light or fluorescence microscopy. The fluorescence microscopy occupies a significant place in biological research due to its ability to address various biological question at subcellular level of a biological sample. One of the more recent developments in fluorescence ...
Dissertation
Characterization of DnmA, the Dnmt2 homolog in Dictyostelium discoideum
(2010-08-24)
Eukaryotic DNA m5C methyltransferases (MTases) play a major role in many epigenetic regulatory processes like genomic imprinting, X-chromosome inactivation, silencing of transposons and gene expression. Members of the two DNA m5C MTase families, Dnmt1 and Dnmt3, are relatively well studied and many details of their biological functions, biochemical properties as well as interaction partners are known. In contrast, the biological functions of the highly conserved Dnmt2 family, which appear to have non-canonical dual ...
Dissertation
Strukturelle und funktionelle Charakterisierung des Dnmt2-Homologs DnmA von Dictyostelium discoideum
(2016-04-08)
In dieser Arbeit wurde die DNA Methyltransferase 2 aus Dictyostelium discoideum strukturell und funktionell untersucht. Sie vermittelt die Methylierung des Cytosin an Position 38 nahe des Anticodons der tRNAAsp (Goll et al., 2006; Müller et al., 2013). Jedoch ist die biologische Funktion dieser Methylierung bis heute nicht hinreichend geklärt.
In der Vergangenheit konnten erhebliche Unterschiede in der in vivo- und in vitro-Methylierungsaktivität von DnmA, dem Dnmt2-Homolog aus D. discoideum, beobachtet werden. So ...
Dissertation
Zelluläre und biochemische Charakterisierung der bifunktionalen Methyltransferase Dnmt2
(2014-09-12)
Seit der Entdeckung der Methyltransferase 2 als hoch konserviertes und weit verbreitetes Enzym sind zahlreiche Versuche zur vollständigen Charakterisierung erfolgt. Dabei ist die biologische Funktion des Proteins ein permanent umstrittener Punkt. In dieser Arbeit wird dnmA als sensitiver Oszillator bezüglich des Zellzyklus und weiterer Einflüsse gezeigt. Insgesamt liegt der Hauptfokus auf der Untersuchung der in vivo Charakterisierung des Gens, der endogenen subzellulären Verteilung, sowie der physiologischen Aufgaben ...
Dissertation
Artificial micro RNA system in Dictyostelium discoideum
(2011-01-12)
Dictyostelium discoideum is a social amoeba that serves as a model system for RNA interference and related mechanisms. Its position between plants and animals enables evolutionary snapshot of mechanisms and protein machinery involved in investigated subjects. MiRNAs are small regulatory RNAs that are evolutionary conserved and present in animals, plants, viruses and some prokaryotes. They have roles in development, cell growth and differentiation, apoptosis and their miss-regulation is associated with many diseases ...
Dissertation
Funktion und Biogenese kleiner RNAs in Dictyostelium discoideum
(2016-01-19)
RNA mediated gene silencing pathways are highly conserved among eukaryotes and they have been well investigated in animals and in plants. Longer dsRNA molecules trigger the silencing pathways: RNase III proteins and their dsRNA binding protein (dsRBP) partners recognize those molecules as a substrate and process 21 nucleotide long microRNAs (miRNAs) or small interfering RNAs (siRNAs). Some organisms encode RNA dependent RNA polymerases (RdRPs), which are able to expand the pool of existing siRNAs. Argonaute proteins ...