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dc.contributor.advisorSpagnolo, Lauraen
dc.contributor.advisorBoettcher, Bettinaen
dc.contributor.authorCannone, Giuseppeen
dc.date.accessioned2016-10-28T13:42:27Z
dc.date.available2016-10-28T13:42:27Z
dc.date.issued2015-06-29
dc.identifier.urihttp://hdl.handle.net/1842/17070
dc.description.abstractPrevious studies suggest a degree of homology between eukaryotic replication, transcription and translation proteins and archaeal ones. Hence, Archaea are considered a simplified model for understanding the complex molecular machinery involved in eukaryotic DNA metabolism. DNA replication in eukaryotic cells is widely studied. In recent years, DNA replication studies expanded on the archaeal DNA replication machinery. P. abyssi was the first archaeon whose genome was fully sequenced. Genome sequencing and comparative genomics have highlighted an MCM-like protein in P. abyssi. In this study, I report the biochemical and structural characterisation of PabMCM. PabMCM is explored as model for understanding more complex eukaryotic MCM proteins and unravelling the biochemical mechanism by which MCM proteins release their helicase activity. The crenarchaeon Sulfolobus solfataricus possesses a simplified toolset for DNA replication compared to Eukaryotes. In particular, S. solfataricus has a subset of the eukaryotic Okazaki fragment maturation factors, among which there are a heterotrimeric DNA sliding clamp, (the proliferating cell nuclear antigen, PCNA), the DNA polymerase B1 (PolB1), the flap endonuclease (Fen1) and the ATP-dependent DNA ligase I (LigI). PCNA functions as a scaffold with each subunit having a specific binding affinity for each of the factors involved in Okazaki fragment maturation. Here, the 3D reconstruction of PCNA in complex with the Okazaki fragment maturation proteins PolB1, LigI and Fen1 is reported.en
dc.contributor.sponsorotheren
dc.language.isoen
dc.publisherThe University of Edinburghen
dc.relation.hasversionZhang, J., Rouillon, C., Kerou, M., Reeks, J., Brugger, K., Graham, S., Reimann, J., Cannone, G., Liu, H., Albers, S.V. et al. (2012) Structure and mechanism of the CMR complex for CRISPR-mediated antiviral immunity. Mol. Cell 45, 303–313en
dc.relation.hasversionRouillon, C., Zhou, M., Zhang, J., Politis, A., Beilsten, V., Cannone, G., Graham, S., Robinson, C., Spagnolo, L. and White, M. (2013) Structure of the CRISPR surveillance complex CSM reveals key similarities with Cascade. Mol. Cell 52, 124–134en
dc.subjecteukaryotic DNA metabolismen
dc.subjecteukaryotic MCM proteinsen
dc.subjectPCNAen
dc.subjectOkazaki fragment maturationen
dc.titleStructural investigation of the archaeal replicative machinery by electron microscopy and digital image processingen
dc.typeThesis or Dissertationen
dc.type.qualificationlevelDoctoralen
dc.type.qualificationnamePhD Doctor of Philosophyen
dc.rights.embargodate2100-12-31
dcterms.accessRightsRestricted Accessen


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