Use this URL to cite or link to this record in EThOS: https://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.799050
Title: Investigation into the role of LSH ATPase in chromatin remodelling
Author: Gukova, Alina
ISNI:       0000 0004 8509 5431
Awarding Body: University of Edinburgh
Current Institution: University of Edinburgh
Date of Award: 2020
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Abstract:
Chromatin remodelling is a crucial nuclear process affecting replication, transcription and repair. Global reduction of DNA methylation is observed in Immunodeficiency-Centromeric Instability-Facial Anomaly (ICF) syndrome. Several proteins were found to be mutated in patients diagnosed with ICF, among them are LSH and CDCA7. LSH is a chromatin remodeller bearing homology to the members of Sf2 remodelling family. A point mutation in its ATPase lobe was identified in ICF. CDCA7 is a zinc finger protein that was recently found to be crucial for nucleosome remodelling activity of LSH. Several point mutations in its zinc finger domain were described in ICF patients. In vitro and in vivo studies have shown that LSH-/- phenotype demonstrates reduction of global DNA methylation, implying that chromatin remodelling LSH functions may be required for the efficient methyltransferase activity, linking this finding to ICF phenotype. Here, the LSH-mononucleosome interaction was explored in vitro using bioinformatics, biochemical, biophysical and structural techniques. LSH purification was further optimised, achieving near 100% purity, which is a useful improvement for any potential structural studies. LSH has been found to interact with the mononucleosome in vitro and no DNA linker was required for this interaction, indicating that LSH binds to the nuclesomal core through its ATPase domain. Qualitatively estimated Kd for this interaction was in nanomolar region, which did not translate into complex detection during size exclusion chromatography. CDCA7 was expressed in insect cell system and semi-purified, however, high nucleic acid presence in the final protein sample precluded any potential studies of CDCA7 interaction with chromatin. Homology and ab initio modelling for LSH and CDCA7, respectively, indicated that LSH is likely to bind the superhelical location 2 (SHL2), however, the exact location of CDCA7 and its interaction with LSH will have to be elucidated in further experimental work.
Supervisor: Richardson, Julia ; Voigt, Philipp Sponsor: Not available
Qualification Name: Thesis (Ph.D.) Qualification Level: Doctoral
EThOS ID: uk.bl.ethos.799050  DOI:
Keywords: histones ; chromatin ; CDCA7 ; ICF syndrome ; LSH
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