Thesis (Ph.D.)--McMaster University, 2003. / Advisor: A. K. Grover. Also available via World Wide Web.

Thesis (Ph.D. in Molecular Biology) -- University of Colorado at Denver and Health Sciences Center, 2005. / Typescript. Includes bibliographical references (leaves 130-152). Free to UCDHSC affiliates. Online version available via ProQuest Digital Dissertations;

Würzburg, Univ., Diss., 2002. / Dateien im PDF-Format

University, Diss., 2004--Heidelberg.

Six ribosomal protein genes from the sulfur dependent extreme thermophilic archaebacterium Sulfolobus solfataricus were cloned and sequenced. Four of these genes code for proteins that are equivalent to ribosomal proteins L11, L1, L10 and L12 in Escherichia coli. The other two genes code for proteins that have no equivalent in the eubacteria. The product of one of these genes was found to be equivalent to ribosomal proteins L46 from yeast (Leer et al. 1985a) and L39 from rat liver (Lin et al. 1984), while the product of the other gene shows no sequence similarity to any of the ribosomal proteins present in the data base. In Sulfolobus, the genes that code for ribosomal proteins L11, L1, L10 and L12 are organized in the same order as in Escherichia coli, that is 5' L11, L1, L10, L12 3'. The major transcript from this gene cluster was found to be a 2.5 Kb mRNA that contains the four genes. A less abundant transcript containing only the L10 and L12 gene was also detected. Upstream of the transcription initiation sites, sequences that match the consensus sequence for archaebacterial promoters (TTTAT/AA) were found. Transcription termination sites were located within or after pyrimidine rich regions. Three of the ribosomal protein genes start with unusual initiation codons, GTG in the case of the L1 and L10 genes and TTG in the case of the L11 gene. Putative Shine Dalgarno sequences, complementary to the 3' end of Sulfolobus 16S rRNA, were detected in the region surrounding the initiation codon. In some cases (L1 and L10 genes), the initiation codon was found to be part of this sequence. Sequence comparison of the ribosomal proteins from Sulfolobus with those from other organisms, revealed that the Sulfolobus sequences are closer to those from other archaebacteria, thus supporting the existence of the archaebacterial kingdom. Comparison of the sequences of the L10 and L12 proteins from the three kingdoms revealed that the archaebacterial sequences are closer to the eukaryotes. / Graduate

Most eukaryotic mRNAs have a sequence of polyadenylic acid [poly(A)] at their 3'-termini. Although it has been almost two decades since the discovery of these poly(A) tracts, their function(s) have yet to be clarified. Earlier results from our laboratory led us to propose that poly(A) has a role in translation. More specifically, we proposed that an interaction of the cytoplasmic poly(A)-binding protein (PABP) with a critical minimum length of poly(A) facilitates the initiation of translation of poly(A)+, but not poly(A)-, mRNAs. The results of several different experimental approaches have provided evidence which indirectly supports this hypothesis. These results include: 1) the correlation of specific changes in mRNA poly(A) tail length with translational efficiency in vivo and in vitro; 2) correlations between the abundance and stability of PABPs and the rate of translational initiation in vivo and in vitro; and 3) the demonstration that exogenous poly(A) is a potent and specific inhibitor of the in vitro translation of poly(A)+, but not poly(A)-mRNAs. To evaluate the hypothesis that the 3'-poly(A) tract of mRNA plays a role in translational initiation, we have constructed derivatives of pSP65 which direct the in vitro synthesis of mRNAs with different poly(A) tail lengths and compared, in reticulocyte extracts, the relative efficiencies with which such mRNAs are translated, degraded, recruited into polysomes, and assembled into mRNPs or intermediates in the translational initiation pathway. Relative to mRNAs which are polyadenylated, we find that poly(A)- mRNAs have a reduced translational capacity which is not due to an increase in their decay rates, but is attributable to a reduction in their efficiency of recruitment into polysomes. The defect in poly(A)- mRNAs affects a late step in translational initiation, is distinct from the phenotype associated with cap-deficient mRNAs, and results in a reduced ability to form 80S initiation complexes. Moreover, poly(A) added in trans inhibits translation from capped poly(A)+ mRNAs, but stimulates translation from capped poly(A)- mRNAs. We suggest that poly(A) is the formal equivalent of a transcriptional enhancer, i.e., that poly(A)-binding protein (PABP) bound at the 3'-end of mRNA may facilitate the binding of an initiation factor or ribosomal subunit at the mRNA 5'-end.

Abstract Prostate cancer is the most common solid tumor among men in Western industrialized countries. A major problem in prostate cancer treatment is the development of androgen-independence, as androgen-deprivation therapy is the basic therapy for the disease. Molecular mechanisms behind prostate cancer and androgen-independent growth development are poorly known. In this study, subtractive hybridization was used for the generation of a cDNA library specific for prostate cancer. Analysis of the cDNA library revealed over-expression of several ribosomal proteins namely L4, L5, L7a, L23a, L30, L37, S14 and S18, in prostate cancer cell lines. Over-expression of L7a and L37 was also confirmed in prostate cancer tissue samples. Further, cDNA array was used in order to examine differentially expressed genes in androgen-dependent and androgen-independent prostate cancer cell line LNCaP. Monoamine oxidase A, an Expressed Sequence Tag (EST) similar to rat P044, and EST AA412049 were highly over-expressed in androgen-dependent LNCaP cells. Tissue-type plasminogen activator, interferon-inducible protein p78 (MxB), an EST similar to galectin-1, follistatin, fatty acid-binding protein 5, EST AA609749, annexin I, the interferon-inducible gene 1-8U and phospholipase D1 were highly over-expressed in androgen-independent LNCaP cells. The EST similar to rat P044, the EST similar to galectin-1, follistatin, annexin I and the interferon-inducible gene 1-8U were also expressed in benign prostatic hyperplasia tissue. The Y-linked ribosomal protein S4, Mat-8, and EST AA307912 were highly expressed in benign prostatic hyperplasia tissue. In situ hybridization of mouse embryos and adult mouse tissues revealed the expression of TMPRSS2 in the epithelium throughout the gastrointestinal, urogenital and respiratory tracts during development. In human multiple tissue RNA dot blot, the highest level of expression was detected in prostate, and lower levels in colon, stomach and salivary gland. TMPRSS2 transcript levels were significantly higher in prostate cancer tissue between benign and malignant epithelium of prostate cancer patients with untreated disease. Similarly, in poorly differentiated adenocarcinomas, expression in malignant tissue was significantly higher. Enzymatic mutation detection and direct sequencing of TMPRSS2 coding region revealed only one deletion in aggressive disease among 9 non-aggressive and 9 aggressive prostate cancer samples. No other mutations were found. Detected 7-base pair deletion leads to premature stop codon and disruption of serine protease substrate binding and catalytic active site. We cloned several potential genes whose expression is changed during prostate cancer initiation or progression. These genes may serve as prostate cancer markers, and further studies are needed to clarify the expression of these proteins during the disease.

Doctorat en Sciences / info:eu-repo/semantics/nonPublished

Dry Pea or field pea (Pisum sativum L.) is an important cool season legume crop grown in the United States. Field peas are vulnerable to many diseases of which, soil borne diseases including wilt and root rot are of major economic importance and can cause significant reduction in yield. There is a dearth of satisfactory methods for control of root rot and no varieties with complete resistance to Fusarium root rot are currently available. Root rot disease was found to be prevalent in all the major pea growing counties of North Dakota surveyed in 2004, 2005, 2010 and 2011. Fusarium species were the most frequently isolated fungal species from the infected pea roots of which, F. oxysporum and F. avenaceum were the most common. 21 Field pea varieties were screened for resistance against F. avenaceum and F. solani f. sp. pisi, the Fusarium species traditionally associated with root rots of field pea in growth chamber experiments and field trials. Low levels of resistance were detected in a few cultivars but no variety was found to be completely resistant to any of the pathogens tested. Efficiency of precipitated calcium carbonate (PCC) in controlling Fusarium species most commonly associated with root rots was evaluated under in vitro and field conditions. Significant reduction in spore production, spore germination, and dry mycelial weight of Fusarium spp. were detected on PCC amended media in laboratory studies. In greenhouse and field experiments significant reduction in root rot disease severity was observed with PCC application compared to control. Fungal gene expression in artificially infected field pea roots and F. graminearum grown in culture was assessed using the Illumina mRNA-Seq technology. A total of 613 F. graminearum genes were found to be differentially expressed in planta on pea. Functional classes associated with amino acid metabolism, nitrogen metabolism, extracellular polysaccharide degradation, detoxification by degradation and defense related proteins were found to be significantly enriched in the up-regulated gene set as determined using FunCatDB. Expression of four up-regulated genes was confirmed by RT-PCR to validate the inferences from the sequencing results.

Workshop "Netz- und Service-Infrastrukturen" Der Vortrag beleuchtete den Gedanken einer Instant-Messenger Infrastruktur für die Uni und den sich daraus ergebenden Möglichkeiten, Problemen sowie offenen Fragen.