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Characterization of Spiroplasma citri Fibril:Insights into polymerization interface and interaction with other cytoskeletal proteins

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dc.contributor.advisor PANANGHAT, GAYATHRI
dc.contributor.author BAPAT, MRINMAYEE
dc.date.accessioned 2024-03-04T09:28:10Z
dc.date.available 2024-03-04T09:28:10Z
dc.date.issued 2024-03
dc.identifier.citation 152 en_US
dc.identifier.uri http://dr.iiserpune.ac.in:8080/xmlui/handle/123456789/8575
dc.description.abstract Spiroplasma is a cell-wall less helical bacterium. Electron microscopy studies show the presence of a flat, monolayered and membrane-bound ribbon-like cytoskeleton structure which consists of Fibril and 5 MreB paralogs. In the absence of any outer appendages, the cytoskeletal ribbon is thought to be responsible for maintaining the cell shape and motility of this organism. My Ph.D. work aims to characterize Fibril and MreBs using structural and biochemical approaches to understand the mechanism by which these proteins contribute to cell shape and motility. For characterization of Fibril, based on the sequence homology of the N-terminal half of the protein to 5’- methylthioadenosine nucleosidase (MTAN), nucleosidase activity was checked for Fibril. We observed that in spite of similar domain architecture, Fibril does not exhibit any nucleosidase activity. Fibril protein forms constitutive filaments. In absence of any information regarding the filament interface, we incorporated EGFP as a fluorescent tag at the loop regions based on the sequence based secondary structure prediction. We found that despite the EGFP insertions at multiple positions, Fibril readily forms filaments and might not be stable as a monomer. Earlier studies have shown that MreB5 interacts with lipids and Fibril. Our results show that Fibril and MreB5 interact in a nucleotide dependent manner similar to MreB5- liposome interaction. Our findings reveal that Fibril does not possess any enzymatic activity or ligand-dependent polymerization and that it acts as a scaffold for cell shape maintenance which is regulated by MreB5 filament dynamics in Spiroplasma. For biochemical characterization like polymerization dynamics, ATP hydrolysis and checking interaction with other MreBs and Fibril, purification of MreB1 was initiated. Purification trials of MreB1 are also discussed in the thesis. en_US
dc.description.sponsorship IISER PUNE DBT PROJECT CODE:30818415 SERB PROJECT CODE: 31223647 ESCMID SERB TRAVEL GRANT: ITS/2023/005438 INFOSYS:IISER P/InfyFnd/Trv/267 Carl Storm International Diversity (CSID) Award Department of Science and Technology (DST) INSPIRE Faculty Fellowship (IFA12/LSBM-52) Innovative Young Biotechnologist Award (BT/07/IYBA/2013) Department of Biotechnology Membrane Structural Biology Program (BT/PR28833/BRB/10/1705/2018) en_US
dc.language.iso en en_US
dc.subject Research Subject Categories::NATURAL SCIENCES en_US
dc.title Characterization of Spiroplasma citri Fibril:Insights into polymerization interface and interaction with other cytoskeletal proteins en_US
dc.type Thesis en_US
dc.description.embargo 1 Year en_US
dc.type.degree Int.Ph.D en_US
dc.contributor.department Dept. of Biology en_US
dc.contributor.registration 20152005 en_US


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  • PhD THESES [590]
    Thesis submitted to IISER Pune in partial fulfilment of the requirements for the degree of Doctor of Philosophy

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