The treble clef binding motif of the zinc finger metalloprotein utilizes N2S2 binding sites. Whereas other N2S2 metalloproteins function in catalytic roles, zinc fingers serve mostly a structural element, although there has been some evidence that the zinc finger protein can interact with exogenous metal ions in aggregate formation or ion exchange. The work presented within has been aimed at precedents for both of the latter in Zn2+. The use of zinc and cadmium dithiolate complexes as mono- and bidentate S-donor ligands to tungsten carbonyl complexes was explored and the ability of zinc and cadmium complexes to stably bind to W(CO)x (x = 4 and 5) was established. The reactivity of thiolate sulfurs within the bimetallic complexes was examined, gaining an understanding of zinc and cadmium N2S2. The characteristics of these complexes were examined via IR, UV-vis, elemental analysis, and x-ray crystallography spectroscopy. The ability of zinc to act as a scaffold for the synthesis of bisacetylbme-dach in the production and subsequent transfer of the same ligand to exogenous metal ion sources was investigated. Cu2+ and Cd2+ analogs to the Zn-1’-Ac2 were synthesized and their properties investigated with IR, elemental analysis, and UV-vis spectroscopy.

Thesis (Ph. D.)--School of Biological Sciences. University of Missouri--Kansas City, 2008. / "A dissertation in cell biology and biophysics and molecular biology and biochemistry." Advisor: John H. Laity. Typescript. Vita. Title from "catalog record" of the print edition Description based on contents viewed Sept.12, 2008. Includes bibliographical references (leaves 148-166). Online version of the print edition.

Thesis (Ph. D.)--Ohio State University, 2004. / Document formatted into pages; contains xvi, 162 p.; also includes grpahics (some col.). Includes bibliographical references. Abstract available online via OhioLINK's ETD Center; full text release delayed at author's request until 2006 May 4.

AreA is a transcription activation protein regulating over 100 genes in Aspergillus nidulans. It is a member of the 4 cysteine zinc finger family, with significant sequence homology in the zinc finger domain with related proteins. As with similar proteins, the zinc finger domain has been identified as the DNA binding motif. Other regions of the protein do not appear to playa role in directly binding DNA. A truncated form of AreA, known as the minimal zinc finger protein, containing the zinc finger domain alone, has been cloned and over-expressed in this study. This domain is sufficient to bind DNA specifically, but weakly. However, the addition of a further 30 amino acids, C-terminal to the minimal zinc finger, containing a highly basic tail is shown to increase the specific binding affinity. Other forms of AreA have been characterised and do not significantly increase the affinity of DNA binding. Identification of a consensus binding sequence by SELEX has also demonstrated that the minimal zinc finger is sufficient to specifically recognise the core sequence GA T A but suggests a degree of tolerance for TAT A. Addition of further regions of the protein do not extend the limits of the recognition sequence or change the consensus sequence. The SELEX experiments did not give any evidence of AreA functioning as a dimer, through proteinprotein interactions. Structural studies of the truncated forms of AreA, by circular dichroism, have suggested the formation of secondary structures for the zinc finger motif similar to those of other proteins and in agreement with the published NMR structure of the AreA zinc finger bound to DNA (Starich et ai., 1998a).

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Thesis (Ph. D.)--University of Texas at Austin, 2002. / Vita. Includes bibliographical references. Available also from UMI Company.

Le principal défi des techniques de transfert de gènes est de garantir l’expression du gène d’intérêt tout en assurant l’innocuité des cellules génétiquement modifiées. La plupart des systèmes d’intégration, dérivés des transposons, assurent une intégration aléatoire au sein du génome de la cellule. L’enjeu de ce travail est de développer des outils permettant de cibler l’intégration du transgène au niveau d’un locus choisi dans le but d’améliorer la biosécurité. L’étude s’est portée sur l’utilisation des protéines à ZFD (Zinc Finger Domain) pour leur aptitude à être conçues à façon, in silico, en utilisant les nombreuses ressources disponibles sur Internet. Pour comparer, les propriétés de deux domaines de liaison, NterR2P, provenant de deux rétrotransposons R2 de type non-LTR, ont été étudiées pour leur capacité naturelle à reconnaître spécifiquement une région de 100 pb située dans l’ADN ribosomique 28S. Les résultats obtenus ont montré que les domaines NterR2P reconnaissent spécifiquement leur ADN cible avec une forte affinité de liaison. Deux protéines de fusion, utilisant le domaine NterR2P, ont ensuite été synthétisées dans le but d’intégrer le transgène au niveau de l’ADNr en utilisant le transposon Sleeping Beauty. L’idée originale de ce travail est de réaliser l’intégration du transgène via le ciblage indirect du transposon, ou de la transposase sans que celle-ci ne soit modifiée. L’impact de ces systèmes de ciblage sur les cellules nécessite de réexaminer une telle stratégie. / The main challenge of gene transfer technologies is to maintain and to sustain transgene expression and to confer innocuity on the genetically-modified cells. Most integration systems, derived from transposons, integrate randomly within the genome of cells. The issue of this work is to develop tools to target transgene integrations in a selected locus in order to improve biosecurity. This study consists in using ZFD (Zinc Finger Domain) proteins for their capability to be in silico synthesize, in using many bioinformatic sites. For compare, the properties of two DNA binding domains (DBD), NterR2P, originating from the endonucleases encoded by R2 non-LTR retrotransposons, are able to bind specifically within a 100-bp region of the 28S rRNA genes. The results show that NterR2P DBDs specifically recognize their DNA target with high affinity. Two fusion proteins, using NterR2P DBD, are synthesized in order to integrate the transgene within rDNA, using Sleeping Beauty transposon. The original idea of this work is to realize transgene integration via indirect targeting of transposon, or transposase without its modification. The use of such a targeting system will have to be extensively studied to determine its impacts on cells before it can be considered as safe for use.

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