An investigation into novel coiled-coil proteins and their roles within the polar growth mechanism.

Salaymeh, Jude Khaled (2023) An investigation into novel coiled-coil proteins and their roles within the polar growth mechanism. Masters thesis, University of East Anglia.

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Abstract

Polar growth is a largely understudied mechanism of bacterial growth found in different pathogenic bacterial species. The link between polar growth, peptidoglycan synthesis, and eventual cell division remains unclear. Streptomyces coelicolor is a model organism, ideal for monitoring the unusual morphological changes, and further investigating polar growth machinery. Using fluorescent d-amino acids, this study mapped peptidoglycan synthesis sites during different life stages of S. coelicolor, to capitalise on the morphological changes and shifts undergone during development. This study highlighted sites of active and dormant peptidoglycan synthesis throughout the Streptomyces life cycle. The mediation of polar growth within S. coelicolor is largely dependent on the Tip-Organising Centre (TIPOC). The intricate molecular mechanisms between TIPOC components are not fully understood. This study aimed to utilise bacterial two hybrid assays to further characterise a newly identified protein, Dia, within TIPOC. The interactions between Dia and chromosome organisation proteins, as well as cell division proteins, were also investigated. The study found evidence to suggest the involvement of Dia with TIPOC, and further involvement in stages of early chromosome organisation. This study indicates that Dia may not be involved within the cell division of S. coelicolor. Using cloning vectors, this study attempted to complement a S. coelicolor scy deletion mutant with an Alpha-proteobacterial gene, lcy, which is hypothesised to implicate the polar growth mechanism of Labrenzia aggregata. Lcy was not complemented in this study; but future testing methods were explored that could further identify a potential homology between Lcy and the vital TIPOC component, Scy. Future complementation attempts could involve the use of a codon optimised protein, a fluorescently tagged complemented Lcy, or reverse complementation of scy and other TIPOC components into L. aggregata.

Item Type:	Thesis (Masters)
Faculty \ School:	Faculty of Science > School of Biological Sciences
Depositing User:	Chris White
Date Deposited:	18 Apr 2024 13:19
Last Modified:	18 Apr 2024 13:19
URI:	https://ueaeprints.uea.ac.uk/id/eprint/94963
DOI:

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