@phdthesis{Scholl2015,
  author    = {Scholl, Christina},
  title     = {Cellular and molecular mechanisms contributing to behavioral transitions and learning in the honeybee},
  url       = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-115527},
  school      = {Universit{\"a}t W{\"u}rzburg},
  year      = {2015},
  abstract  = {The honeybee Apis mellifera is a social insect well known for its complex behavior and the ability to learn tasks associated with central place foraging, such as visual navigation or to learn and remember odor-reward associations. Although its brain is smaller than 1mm² with only 8.2 x 105 neurons compared to ~ 20 x 109 in humans, bees still show amazing social, cognitive and learning skills. They express an age - related division of labor with nurse bees staying inside the hive and performing tasks like caring for the brood or cleaning, and foragers who collect food and water outside the hive. This challenges foragers with new responsibilities like sophisticated navigation skills to find and remember food sources, drastic changes in the sensory environment and to communicate new information to other bees. Associated with this plasticity of the behavior, the brain and especially the mushroom bodies (MBs) - sensory integration and association centers involved in learning and memory formation - undergo massive structural and functional neuronal alterations. Related to this background my thesis on one hand focuses on neuronal plasticity and underlying molecular mechanisms in the MBs that accompany the nurse - forager transition. In the first part I investigated an endogenous and an internal factor that may contribute to the nurse - forager phenotype plasticity and the correlating changes in neuronal network in the MBs: sensory exposure (light) and juvenile hormone (JH). Young bees were precociously exposed to light and subsequently synaptic complexes (microglomeruli, MG) in the MBs or respectively hemolymph juvenile hormone (JH) levels were quantified. The results show that light input indeed triggered a significant decrease in MG density, and mass spectrometry JH detection revealed an increase in JH titer. Interestingly light stimulation in young bees (presumably nurse bees) triggered changes in MG density and JH levels comparable to natural foragers. This indicates that both sensory stimuli as well as the endocrine system may play a part in preparing bees for the behavioral transition to foraging. Considering a connection between the JH levels and synaptic remodeling I used gene knockdown to disturb JH pathways and artificially increase the JH level. Even though the knockdown was successful, the results show that MG densities remained unchanged, showing no direct effect of JH on synaptic restructuring. To find a potential mediator of structural synaptic plasticity I focused on the calcium-calmodulin-dependent protein kinase II (CaMKII) in the second part of my thesis. CaMKII is a protein known to be involved in neuronal and behavioral plasticity and also plays an important part in structural plasticity reorganizing synapses. Therefore it is an interesting candidate for molecular mechanisms underlying MG reorganization in the MBs in the honeybee. Corresponding to the high abundance of CaMKII in the learning center in vertebrates (hippocampus), CaMKII was shown to be enriched in the MBs of the honeybee. Here I first investigated the function of CaMKII in learning and memory formation as from vertebrate work CaMKII is known to be associated with the strengthening of synaptic connections inducing long term potentiation and memory formation. The experimental approach included manipulating CaMKII function using 2 different inhibitors and a specific siRNA to create a CaMKII knockdown phenotype. Afterwards bees were subjected to classical olfactory conditioning which is known to induce stable long-term memory. All bees showed normal learning curves and an intact memory acquisition, short-term and mid-term memory (1 hour retention). However, in all cases long-term memory formation was significantly disrupted (24 and 72 hour retention). These results suggests the necessity of functional CaMKII in the MBs for the induction of both early and late phases of long-term memory in honeybees. The neuronal and molecular bases underlying long-term memory and the resulting plasticity in behavior is key to understanding higher brain function and phenotype plasticity. In this context CaMKII may be an important mediator inducing structural synaptic and neuronal changes in the MB synaptic network.},
  subject      = {Biene},
  language  = {en}
}
@phdthesis{Nuwal2010,
  author    = {Nuwal, Tulip},
  title     = {Characterization of Synapsin, Tubulin-Binding Chaperone E-like, And Their Putative Interactions With Synapse Associated Protein Of 47 kDa In Drosophila melanogaster},
  url       = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-51683},
  school      = {Universit{\"a}t W{\"u}rzburg},
  year      = {2010},
  abstract  = {In this thesis we have used Drosophila melanogaster as a model organism to investigate proteins and their putative interacting partners that are directly or indirectly involved in the release of neurotransmitters at the synapse. We have used molecular techniques to investigate conserved synaptic proteins, synapsin and synapse associated protein of 47 kD (SAP47), and a putative interaction partner of SAP47, tubulin binding chaperone E-like (TBCEL). SAP47 and synapsins are highly conserved synaptic vesicle associated proteins in Drosophila melanogaster. To further investigate the role and function of Sap47 and Syn genes, we had earlier generated the null mutants by P-element mutagenesis (Funk et al., 2004; Godenschwege et al., 2004). Western blots and ELISA of brain homogenates from Sap47156 null mutants showed the presence of up-regulated phospho-synapsin in comparison to wild-type (CS) and the presence of up-regulated phospho-synapsin was partially abolished when a pan-neuronal rescue of SAP47 was performed by the Gal4- UAS technique. Thus, the results suggest a qualitative and quantitative modulation of synapsin by SAP47. At the transcript level, we did not observe any difference in content of Syn transcript in Sap47156 and wild-type CS flies. The question of a direct molecular interaction between SAP47 and synapsin was investigated by co-immunoprecipitation (Co-IP) experiments and we did not find any stable interactions under the several IP conditions we tested. The possibility of Sap47 as a modifier of Syn at the genetic level was investigated by generating and testing homozygous double null mutants of Sap47 and Syn. The Syn97, Sap47156 double mutants are viable but have a reduced life span and decreased locomotion when compared to CS. In 2D-PAGE analysis of synapsins we identified trains of spots corresponding to synapsins, suggesting that synapsin has several isoforms and each one of them is posttranslationally modified. In an analysis by Blue native-SDS-PAGE (BN-SDS-2D- PAGE) and Western blot we observed synapsin and SAP47 signals to be present at 700-900 kDa and 200-250 kDa, respectively, suggesting that they are part of large but different complexes. We also report the possibility of Drosophila synapsin forming homo- and heteromultimers, which has also been reported for synapsins of vertebrates. In parallel to the above experiments, phosphorylation of synapsins in Drosophila was studied by IP techniques followed by 1D-SDS gel electrophoresis and mass spectrometry (in collaboration with S. Heo and G. Lubec). We identified and verified 5 unique phosphorylation sites in Drosophila synapsin from our MS analysis. Apart from phosphorylation modifications we identified several other PTMs which have not been verified. The significance of these phosphorylations and other identified PTMs needs to be investigated further and their implications for synapsin function and Drosophila behavior has to be elucidated by further experiments. In a collaborative project with S. Kneitz and N. Nuwal, we investigated the effects of Sap156 and Syn97 mutations by performing a whole Drosophila transcriptome microarray analysis of the individual null mutants and the double mutants (V2 and V3). We obtained several candidates which were significantly altered in the mutants. These genes need to be investigated further to elucidate their interactions with Sap47 and Syn. In another project, we investigated the role and function of Drosophila tubulin- binding chaperone E-like (Tbcel, CG12214). The TBCEL protein has high homology to vertebrate TBCE-like (or E-like) which has high sequence similarity to tubulin-binding chaperone E (TBCE) (hence the name TBCE-Like). We generated an anti-TBCEL polyclonal antiserum (in collaboration with G. Krohne). According to flybase, the Tbcel gene has only one exon and codes for two different transcripts by alternative transcription start sites. The longer transcript RB is present only in males whereas the shorter transcript RA is present only in females. In order to study the gene function we performed P- element jump-out mutagenesis to generate deletion mutants. We used the NP4786 (NP) stock which has a P(GawB) insertion in the 5' UTR of the Tbcel gene. NP4786 flies are homozygous lethal due to a second-site lethality as the flies are viable over a deficiency (Df) chromosome (a deletion of genomic region spanning the Tbcel gene and other upstream and downstream genes). We performed the P-element mutagenesis twice. In the first trial we obtained only revertants and the second experiment is still in progress. In the second attempt, jump-out was performed over the deficiency chromosome to prevent homologous chromosome mediated double stranded DNA repair. During the second mutagenesis an insertion stock G18151 became available. These flies had a P-element insertion in the open reading frame (ORF) of the Tbcel gene but was homozygous viable. Western blots of fresh tissue homogenates of NP/Df and G18151 flies probed with anti-TBCEL antiserum showed no TBCEL signal, suggesting that these flies are Tbcel null mutants. We used these flies for further immunohistochemical analyses and found that TBCEL is specifically expressed in the cytoplasm of cyst cells of the testes and is associated with the tubulin of spermatid tails in wild-type CS, whereas in NP/Df and G18151 flies the TBCEL staining in the cyst cells was absent and there was a disruption of actin investment cones. We also found enrichment of TBCEL staining around the actin investment cone. These results are also supported by the observation that the enhancer trap expression of the NP4786 line is localised to the cyst cells, similar to TBCEL expression. Also, male fertility of NP/Df and G18151 flies was tested and they were found to be sterile with few escapers. Thus, these results suggest that TBCEL is involved in Drosophila spermatogenesis with a possible role in the spermatid elongation and individualisation process.},
  subject      = {Taufliege},
  language  = {en}
}
@phdthesis{Ott2013,
  author    = {Ott, Christine Kornelia},
  title     = {Diverse Aspects of the Sorting and Assembly Machinery in Human Mitochondria},
  url       = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-85462},
  school      = {Universit{\"a}t W{\"u}rzburg},
  year      = {2013},
  abstract  = {Mitochondria are organelles of endosymbiotic origin, which play many important roles in eukaryotic cells. Mitochondria are surrounded by two membranes and, considering that most of the mitochondrial proteins are produced in the cytosol, possess import machineries, which transport mitochondria-targeted proteins to their designated location. A special class of outer mitochondrial membrane (OMM) proteins, the β-barrel proteins, require the sorting and assembly machinery (SAM) for their OMM integration. Both mitochondrial β-barrel proteins and the central component of the SAM complex, Sam50, have homologs in gram-negative bacteria. In yeast mitochondria, bacterial β-barrel proteins can be imported and assembled into the OMM. Our group demonstrated that this, however, is not the case for human mitochondria, which import only neisserial β barrel proteins, but not those of Escherichia coli and Salmonella enterica. As a part of this study, I could demonstrate that β-barrel proteins such as Omp85 and PorB of different Neisseria species are targeted to human mitochondria. Interestingly, only proteins belonging to the neisserial Omp85 family were integrated into the OMM, whereas PorB was imported into mitochondria but not assembled. By exchanging parts of homologous neisserial Omp85 and E. coli BamA and, similarly, of neisserial PorB and E. coli OmpC, it could be demonstrated in this work that the mitochondrial import signal of bacterial β barrel proteins cannot be limited to one short linear sequence, but rather secondary structure and protein charge seem to play an important role, as well as specific residues in the last β-strand of Omp85. Omp85 possesses five conserved POTRA domains in its amino-terminal part. This work additionally demonstrated that in human mitochondria, at least two POTRA domains of Omp85 are necessary for membrane integration and functionality of Omp85. In the second part of this work, the influence of Sam50 on the mitochondrial cristae structure was investigated. This work contributed to a study performed by our group in which it was confirmed that Sam50 is present in a high molecular weight complex together with mitofilin, CHCHD3, CHCHD6, DnaJC11, metaxin 1 and metaxin 2. This connection between the inner and outer mitochondrial membrane was shown to be crucial for the maintenance of the mitochondrial cristae structure. In addition, a role of Sam50 in respiratory complex assembly, suggested by a SILAC experiment conducted in our group, could be confirmed by in vitro import studies. An influence of Sam50 not only on respiratory complexes but also on the recently described respiratory complex assembly factor TTC19 was demonstrated. It was shown that TTC19 not only plays a role in complex III assembly as published, but also influences the assembly of respiratory complex IV. Thus, in this part of the work a connection between the OMM protein Sam50 and maintenance of cristae structure, respiratory complex assembly and an assembly factor could be established.},
  subject      = {Mitochondrien},
  language  = {en}
}
@phdthesis{Liu2022,
  author    = {Liu, Ruiqi},
  title     = {Dynamic regulation of the melanocortin 4 receptor system in body weight homeostasis and reproductive maturation in fish},
  doi       = {10.25972/OPUS-20653},
  url       = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-206536},
  school      = {Universit{\"a}t W{\"u}rzburg},
  year      = {2022},
  abstract  = {Puberty is an important period of life with physiological changes to enable animals to reproduce. Xiphophorus fish exhibit polymorphism in body size, puberty timing, and reproductive tactics. These phenotypical polymorphisms are controlled by the Puberty (P) locus. In X. nigrensis and X. multilineatus, the P locus encodes the melanocortin 4 receptor (Mc4r) with high genetic polymorphisms. Mc4r is a member of the melanocortin receptors, belonging to class A G-protein coupled receptors. The Mc4r signaling system consists of Mc4r, the agonist Pomc (precursor of various MSH and of ACTH), the antagonist Agrp and accessory protein Mrap2. In humans, MC4R has a role in energy homeostasis. MC4R and MRAP2 mutations are linked to human obesity but not to puberty. Mc4rs in X. nigrensis and X. multilineatus are present in three allele classes, A, B1 and B2, of which the X-linked A alleles express functional receptors and the male-specific Y-linked B alleles encode defective receptors. Male body sizes are correlated with B allele type and B allele copy numbers. Late-maturing large males carry B alleles in high copy number while early-maturing small males carry B alleles in low copy number or only A alleles. Cell culture co-expression experiments indicated that B alleles may act as dominant negative receptor mutants on A alleles. In this study, the main aim was to biochemically characterize the mechanism of puberty regulation by Mc4r in X. nigrensis and X. multilineatus, whether it is by Mc4r dimerization and/or Mrap2 interaction with Mc4r or other mechanisms. Furthermore, Mc4r in X. hellerii (another swordtail species) and medaka (a model organism phylogenetically close to Xiphophorus) were investigated to understand if the investigated mechanisms are conserved in other species. In medaka, the Mc4r signaling system genes (mc4r, mrap2, pomc, agrp1) are expressed before hatching, with agrp1 being highly upregulated during hatching and first feeding. These genes are mainly expressed in adult brain, and the transcripts of mrap2 co-localize with mc4r indicating a function in modulating Mc4r signaling. Functional comparison between wild-type and mc4r knockout medaka showed that Mc4r knockout does not affect puberty timing but significantly delays hatching due to the retarded embryonic development of knockout medaka. Hence, the Mc4r system in medaka is involved in regulation of growth rather than puberty. In Xiphophorus, expression co-localization of mc4r and mrap2 in X. nigrensis and X. hellerii fish adult brains was characterized by in situ hybridization. In both species, large males exhibit strikingly high expression of mc4r while mrap2 shows similar expression level in the large and small male and female. Differently, X. hellerii has only A-type alleles indicating that the puberty regulation mechanisms evolved independently in Xiphophorus genus. Functional analysis of Mrap2 and Mc4r A/B1/B2 alleles of X. multilineatus showed that increased Mrap2 amounts induce higher cAMP response but EC50 values do not change much upon Mrap2 co-expression with Mc4r (expressing only A allele or A and B1 alleles). A and B1 alleles were expressed higher in large male brains, while B2 alleles were only barely expressed. Mc4r A-B1 cells have lower cAMP production than Mc4r A cells. Together, this indicates a role of Mc4r alleles, but not Mrap2, in puberty onset regulation signaling. Interaction studies by FRET approach evidenced that Mc4r A and B alleles can form heterodimers and homodimers in vitro, but only for a certain fraction of the expressed receptors. Single-molecule colocalization study using super-resolution microscope dSTORM confirmed that only few Mc4r A and B1 receptors co-localized on the membrane. Altogether, the species-specific puberty onset regulation in X. nigrensis and X. multilineatus is linked to the presence of Mc4r B alleles and to some extent to its interaction with A allele gene products. This is reasoned to result in certain levels of cAMP signaling which reaches the dynamic or static threshold to permit late puberty in large males. In summary, puberty onset regulation by dominant negative effect of Mc4r mutant alleles is a special mechanism that is found so far only in X. nigrensis and X. multilineatus. Other Xiphophorus species obviously evolved the same function of the pathway by diverse mechanisms. Mc4r in other fish (medaka) has a role in regulation of growth, reminiscent of its role in energy homeostasis in humans. The results of this study will contribute to better understand the biochemical and physiological functions of the Mc4r system in vertebrates including human.},
  subject      = {Japank{\"a}rpfling},
  language  = {en}
}
@phdthesis{Berkane2005,
  author    = {Berkane, Emir},
  title     = {Etude de l'interaction entre GpJ, une prot{\´e}ine du bact{\´e}riophage Lambda, et LamB, une prot{\´e}ine de la membrane externe des bact{\´e}ries gram-n{\´e}gatives},
  url       = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-12889},
  school      = {Universit{\"a}t W{\"u}rzburg},
  year      = {2005},
  abstract  = {La fixation du bact{\´e}riophage Lambda sur son r{\´e}cepteur cellulaire, LamB, est d{\^u}e {\`a} une prot{\´e}ine de sa queue appel{\´e}e GpJ. Le but des travaux est d'{\´e}tudier l'int{\´e}raction entre le bact{\´e}riophage Lambda et LamB {\`a} travers l'{\´e}tude du complexe entre LamB et GpJ exprim{\´e}e en prot{\´e}ine de fusion. Pour ce faire, deux prot{\´e}ines de fusion sont utilis{\´e}es : MBP-gpJ et HisgpJ. MBP-gpJ est une prot{\´e}ine de fusion entre la Maltose Binding Prot{\´e}ine et l'extr{\^e}mit{\´e} Cterminale de la prot{\´e}ine GpJ (r{\´e}sidu 684 {\`a} 1132), gr{\^a}cieusement fournie par le Pr. Charbit (Paris, France). Gr{\^a}ce {\`a} la Technique du Film Noir (BLM), il a {\´e}t{\´e} permis d'observer que MBP-gpJ, apr{\`e}s expression dans E.coli et purification, int{\´e}ragit gr{\^a}ce au fragment de GpJ avec l'extr{\^e}mit{\´e} extracellulaire de LamB. Cette int{\´e}raction se traduit par un blocage complet et r{\´e}versible des canaux de LamB sauvage, mais {\´e}galement de mutants: LamB de Shigella sonnei, LamB Y118G et LamB D4+D6+D9v. Afin d'obtenir des informations sur la liaison de LamB avec uniquement le fragment de GpJ sans la partie MBP, une autre prot{\´e}ine de fusion a {\´e}t{\´e} r{\´e}alis{\´e}e: His-GpJ. His-gpJ repr{\´e}sente l'extr{\^e}mit{\´e} C-terminale de GpJ (684-1132) en fusion avec un 6×Histidine-tag. Cette prot{\´e}ine est exprim{\´e}e sous forme de corps d'inclusion dans E.coli. Apr{\`e}s purification et renaturation, une prot{\´e}ine de nouveau soluble peut {\^e}tre obtenue. Lors d'exp{\´e}riences de Film Noir, His-gpJ int{\´e}ragit certes avec LamB, mais n'induit pas le blocage des canaux comme pr{\´e}cedemment observ{\´e} apr{\`e}s ajout de MBP-gpJ. En parall{\`e}le, la formation d'un complexe entre His-gpJ et LamB sauvage, ainsi que de mutants a pu {\^e}tre confirm{\´e}e au travers de travaux de SDS-PAGE et d'immunod{\´e}tection par la pr{\´e}sence de bandes de masse mol{\´e}culaire {\´e}lev{\´e}e. L'utilisation de mutants de LamB a par ailleurs permis d'essayer d'identifier la partie de LamB impliqu{\´e}e dans l'interaction avec le fragment C-terminal de GpJ, qui se r{\´e}v{\`e}le {\^e}tre diff{\´e}rente de celle de GpJ dans la queue du bact{\´e}riophage Lambda. Mots cl{\´e}s: bact{\´e}riophage Lambda, gpJ, LamB, technique du film noir (BLM), immunod{\´e}tection.},
  subject      = {Bakteriophage Lambda},
  language  = {en}
}
@phdthesis{Kern2014,
  author    = {Kern, Selina Melanie},
  title     = {Functional characterization of splicing-associated kinases in the blood stages of the malaria parasite Plasmodium falciparum},
  url       = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-115219},
  school      = {Universit{\"a}t W{\"u}rzburg},
  year      = {2014},
  abstract  = {Besides HIV and tuberculosis, malaria still is one of the most devastating infectious diseases especially in developing countries, with Plasmodium falciparum being responsible for the frequently lethal form of malaria tropica. It is a major cause of mortality as well as morbidity, whereby pregnant women and children under the age of five years are most severely affected. Rapidly emerging drug resistances and the lack of an effective and safe vaccine hamper the combat against malaria by chemical and pharmacological regimens, and moreover the poor socio-economic and healthcare conditions in malaria-endemic countries are compromising the extermination of this deadly tropical disease to a large extent. Malaria research is still questing for druggable targets in the parasitic protozoan which pledge to be refractory against evolving resistance-mediating mutations and yet constitute affordable and compliant antimalarial chemotherapeutics. The parasite kinome consists of members that represent most eukaryotic protein kinase groups, but also contains several groups that can not be assigned to conservative ePK groups. Moreover, given the remarkable divergence of plasmodial kinases in respect to the human host kinome and the fact that several plasmodial kinases have been identified that are essential for the intraerythrocytic developmental cycle, these parasite enzymes represent auspicious targets for antimalarial regimens. Despite elaborate investigations on several other ePK groups, merely scant research has been conducted regarding the four identified members of the cyclin-dependent kinase-like kinase (CLK) family, PfCLK-1-4. In other eukaryotes, CLKs are involved in mRNA processing and splicing by means of phosphorylation of serine/arginine-rich (SR) proteins, which are crucial components of the splicing machinery in the alternative splicing pathway. All four PfCLKs are abundantly expressed in asexual parasites and gametocytes, and stage-specific expression profiles of PfCLK-1 and PfCLK-2 exhibited nucleus-associated localization and an association with phosphorylation activity. In the course of this study, PfCLK-3 and PfCLK-4 were functionally characterized by indirect immunofluorescence, Western blot analysis and kinase activity assays. These data confirm that the two kinases are primarily expressed in the nucleus of trophozoites and both kinases possess in vitro phosphorylation activity on physiological substrates. Likewise PfCLK-1 and PfCLK-2, reverse genetic studies exhibited the indispensability of both PfCLKs on the asexual life cycle of P. falciparum, rendering them as potential candidates for antiplasmodial strategies. Moreover, this study was conducted to identify putative SR proteins as substrates of all four PfCLKs. Previous alignments revealed a significant homology of the parasite CLKs to yeast SR protein kinase Sky1p. Kinase activity assays showed in vitro phosphorylation of the yeast Sky1p substrate and SR protein Npl3p by precipitated PfCLKs. In addition, four homologous plasmodial SR proteins were identified that are phosphorylated by PfCLKs in vitro: PfASF-1, PFSRSF12, PfSFRS4 and PfSR-1. All four parasite SR splicing factors are predominantly expressed in the nuclei of trophozoites. For PfCLK-1, a co-localization with the SR proteins was verified. Finally, a library of human and microbial CLK inhibitors and the antiseptic chlorhexidine (CHX) was screened to determine their inhibitory effect on different parasite life cycle stages and on the PfCLKs specifically. Five inhibitors out of 63 compounds from the investigated library were selected that show a moderate inhibition on asexual life cycle stages with IC50 values ranging between approximately 4 and 8 µM. Noteworthy, these inhibitors belong to the substance classes of aminopyrimidines or oxo-β-carbolines. Actually, the antibiotic compound CHX demonstrated an IC50 in the low nanomolar range. Stage-of-inhibition assays revealed that CHX severely affects the formation of schizonts. All of the selected CLKs inhibitors also affect gametocytogenesis as well as gametogenesis, as scrutinized in gametocyte toxicity assays and exflagellation assays, respectively. Kinase activity assays confirm a specific inhibition of CLK-mediated phosphorylation of all four kinases, when the CLK inhibitors are applied on immunoprecipitated PfCLKs. These findings on PfCLK-inhibiting compounds are initial attempts to determine putative antimalarial compounds targeting the PfCLKs. Moreover, these results provide an effective means to generate chemical kinase KOs in order to phenotypically study the role of the PfCLKs especially in splicing events and mRNA metabolism. This approach of functionally characterizing the CLKs in P. falciparum is of particular interest since the malarial spliceosome is still poorly understood and will gain further insight into the parasite splicing machinery.},
  subject      = {Plasmodium falciparum},
  language  = {en}
}
@incollection{AltschmiedSchartl1994,
  author    = {Altschmied, Joachim and Schartl, Manfred},
  title     = {Genetics and molecular biology of tumour formation in Xiphophorus},
  url       = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-69752},
  publisher      = {Universit{\"a}t W{\"u}rzburg},
  year      = {1994},
  abstract  = {No abstract available.},
  subject      = {Schwertk{\"a}rpfling},
  language  = {en}
}
@phdthesis{Wietek2001,
  author    = {Wietek, Irina},
  title     = {Human Interleukin-4 binding protein epitope involved in high-affinity binding of interleukin-4},
  url       = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-3190},
  school      = {Universit{\"a}t W{\"u}rzburg},
  year      = {2001},
  abstract  = {No abstract available},
  subject      = {Mensch},
  language  = {en}
}
@phdthesis{Surrey2020,
  author    = {Surrey, Verena},
  title     = {Identification of affected cellular targets, mechanisms and signaling pathways in a mouse model for spinal muscular atrophy with respiratory distress type 1 (SMARD1)},
  doi       = {10.25972/OPUS-17638},
  url       = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-176386},
  school      = {Universit{\"a}t W{\"u}rzburg},
  year      = {2020},
  abstract  = {Spinal muscular atrophy with respiratory distress type 1 (SMARD1) is a fatal monogenic motoneuron disease in children with unknown etiology caused by mutations in the immunoglobulin μ-binding protein 2 (IGHMBP2) gene coding for DNA/RNA ATPase/helicase. Despite detailed knowledge of the underlying genetic changes, the cellular mechanisms leading to this disease are not well understood. In the Nmd2J ("neuromuscular disorder") mouse, the mouse model for the juvenile form of SMARD1 patients, in which similar pathological features as diaphragmatic paralysis and skeletal muscle atrophy are observed. Ex vivo studies in Nmd2J mice showed that loss of the motor axon precedes atrophy of the gastrocnemius muscle and does not correlate with neurotransmission defects in the motor endplate. The already described independent myogenic anomalies in the diaphragm and heart of the Nmd2J mouse raised the question whether spinal motoneuron degeneration develops cell autonomously. Ighmbp2 is predominantly localized in the cytoplasm and seems to bind to ribosomes and polysomes, suggesting a role in mRNA metabolism. In this Ph.D. thesis, morphological and functional analyses of isolated Ighmbp2-deficient (Ighmbp2-def.) motoneurons were performed to answer the question whether the SMARD1 phenotype results from dysregulation of protein biosynthesis. Ighmbp2-deficient motoneurons show only negligible morphological alterations with respect to a slight increase in axonal branches. This observation is consistent with only minor changes of transcriptome based on RNA sequencing data from Ighmbp2-deficient motoneurons. Only the mRNA of fibroblast growth factor receptor 1 (Fgfr1) showed significant up-regulation in Ighmbp2-deficient motoneurons. Furthermore, no global aberrations at the translational level could be detected using pulsed SILAC (Stable Isotope Labeling by Amino acids in cell culture), AHA (L-azidohomoalanine) labeling and SUnSET (SUrface SEnsing of Translation) methods. However, a reduced β-actin protein level was observed at the growth cones of Ighmbp2-deficient motoneurons, which was accompanied with a reduced level of Imp1 protein, a known β-actin mRNA interactor. Live-cell imaging studies using fluorescence recovery after photobleaching (FRAP) showed translational down-regulation of eGFPmyr-β-actin 3'UTR mRNA in the growth cones and the cell bodies, although the amount of β-actin mRNA and the total protein amount in Ighmbp2-deficient motoneurons showed no aberrations. This compartment-specific reduction of β-actin protein occurred independently of a non-existent direct IGHMBPF2 binding to β-actin mRNA. Fgfr1, which was upregulated on the RNA level, did not show an increased protein amount in Ighmbp2-deficient motoneurons, whereas a reduced amount could be detected. Interestingly, a correlation could be found between the reduced amount of the Imp1 protein and the increased Fgfr1 mRNA, since the IMP1 protein binds the FGFR1 mRNA and thus could influence the transport and translation of FGFR1 mRNA. In summary, all data suggest that Ighmbp2 deficiency leads to a local but modest disturbance of protein biosynthesis, which might contribute to the motoneuron defects of SMARD1.},
  subject      = {Spinale Muskelatrophie},
  language  = {en}
}
@phdthesis{Dagvadorj2016,
  author    = {Dagvadorj, Nergui},
  title     = {Improvement of T-cell response against WT1-overexpressing leukemia by newly developed anti-hDEC205-WT1 antibody fusion proteins},
  url       = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-149098},
  school      = {Universit{\"a}t W{\"u}rzburg},
  year      = {2016},
  abstract  = {Wilms tumor protein 1 (WT1) is a suitable target to develop an immunotherapeutic approach against high risk acute myeloid leukemia (AML), particularly their relapse after allogeneic hematopoietic stem cell transplantation (HSCT). As an intracellular protein traversing between nucleus and cytoplasm, recombinant expression of WT1 is difficult. Therefore, an induction of WT1-specific T-cell responses is mostly based on peptide vaccination as well as dendritic cell (DC) electroporation with mRNA encoding full-length protein to mount WT1-derived peptide variations presented to T cells. Alternatively, the WT1 peptide presentation could be broadened by forcing receptor-mediated endocytosis of DCs. In this study, antibody fusion proteins consisting of an antibody specific to the human DEC205 endocytic receptor and various fragments of WT1 (anti-hDEC205-WT1) were generated for a potential DC-targeted recombinant WT1 vaccine. Anti-hDEC205-WT1 antibody fusion proteins containing full-length or major parts of WT1 were not efficiently expressed and secreted due to their poor solubility and secretory capacity. However, small fragment-containing variants: anti-hDEC205-WT110-35, anti-hDEC205-WT191-138, anti-hDEC205-WT1223-273, and anti-hDEC205-WT1324-371 were obtained in good yields. Since three of these fusion proteins contain the most of the known immunogenic epitopes in their sequences, the anti-hDEC205-WT191-138, anti-hDEC205-WT1223-273, and anti-hDEC205-WT1324-371 were tested for their T-cell stimulatory capacities. Mature monocyte-derived DCs loaded with anti-hDEC205-WT191-138 could induce ex vivo T-cell responses in 12 of 16 blood samples collected from either healthy or HSC transplanted individuals compared to included controls (P < 0.01). Furthermore, these T cells could kill WT1-overexpressing THP-1 leukemia cells in vitro after expansion. In conclusion, alongside proving the difficulty in expression and purification of intracellular WT1 as a vaccine protein, our results from this work introduce an alternative therapeutic vaccine approach to improve an anti-leukemia immune response in the context of allogeneic HSCT and potentially beyond.},
  subject      = {Akute myeloische Leuk{\"a}mie},
  language  = {en}
}